Cell-Free System: A fractionated cell extract that maintains a biological function. A subcellular fraction isolated by ultracentrifugation or other separation techniques must first be isolated so that a process can be studied free from all of the complex side reactions that occur in a cell. The cell-free system is therefore widely used in cell biology. (From Alberts et al., Molecular Biology of the Cell, 2d ed, p166)Protein Biosynthesis: The biosynthesis of PEPTIDES and PROTEINS on RIBOSOMES, directed by MESSENGER RNA, via TRANSFER RNA that is charged with standard proteinogenic AMINO ACIDS.DNA: A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.Cell Line: Established cell cultures that have the potential to propagate indefinitely.Kinetics: The rate dynamics in chemical or physical systems.Molecular Sequence Data: Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.RNA, Messenger: RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm.Transcription, Genetic: The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.Escherichia coli: A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.Cells, Cultured: Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.History, 18th Century: Time period from 1701 through 1800 of the common era.Career Choice: Selection of a type of occupation or profession.History, 19th Century: Time period from 1801 through 1900 of the common era.Career Mobility: The upward or downward mobility in an occupation or the change from one occupation to another.History, 20th Century: Time period from 1901 through 2000 of the common era.Cytokinesis: The process by which the CYTOPLASM of a cell is divided.Cell Biology: The study of the structure, behavior, growth, reproduction, and pathology of cells; and the function and chemistry of cellular components.BostonXenopus: An aquatic genus of the family, Pipidae, occurring in Africa and distinguished by having black horny claws on three inner hind toes.Xenopus laevis: The commonest and widest ranging species of the clawed "frog" (Xenopus) in Africa. This species is used extensively in research. There is now a significant population in California derived from escaped laboratory animals.Xenopus Proteins: Proteins obtained from various species of Xenopus. Included here are proteins from the African clawed frog (XENOPUS LAEVIS). Many of these proteins have been the subject of scientific investigations in the area of MORPHOGENESIS and development.Logic: The science that investigates the principles governing correct or reliable inference and deals with the canons and criteria of validity in thought and demonstration. This system of reasoning is applicable to any branch of knowledge or study. (Random House Unabridged Dictionary, 2d ed & Sippl, Computer Dictionary, 4th ed)Computers, Molecular: Computers whose input, output and state transitions are carried out by biochemical interactions and reactions.Synthetic Biology: A field of biological research combining engineering in the formulation, design, and building (synthesis) of novel biological structures, functions, and systems.Promoter Regions, Genetic: DNA sequences which are recognized (directly or indirectly) and bound by a DNA-dependent RNA polymerase during the initiation of transcription. Highly conserved sequences within the promoter include the Pribnow box in bacteria and the TATA BOX in eukaryotes.Lac Repressors: Bacterial repressor proteins that bind to the LAC OPERON and thereby prevent the synthesis of proteins involved in catabolism of LACTOSE. When lactose levels are high lac repressors undergo an allosteric change that causes their release from the DNA and the resumption of lac operon transcription.Operator Regions, Genetic: The regulatory elements of an OPERON to which activators or repressors bind thereby effecting the transcription of GENES in the operon.Polysaccharides, Bacterial: Polysaccharides found in bacteria and in capsules thereof.PolysaccharidesMast-Cell Sarcoma: A unifocal malignant tumor that consists of atypical pathological MAST CELLS without systemic involvement. It causes local destructive growth in organs other than in skin or bone marrow.Sodium Hydroxide: A highly caustic substance that is used to neutralize acids and make sodium salts. (From Merck Index, 11th ed)O Antigens: The lipopolysaccharide-protein somatic antigens, usually from gram-negative bacteria, important in the serological classification of enteric bacilli. The O-specific chains determine the specificity of the O antigens of a given serotype. O antigens are the immunodominant part of the lipopolysaccharide molecule in the intact bacterial cell. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed)Uridine Diphosphate N-Acetylglucosamine: Serves as the biological precursor of insect chitin, of muramic acid in bacterial cell walls, and of sialic acids in mammalian glycoproteins.Carbohydrate Sequence: The sequence of carbohydrates within POLYSACCHARIDES; GLYCOPROTEINS; and GLYCOLIPIDS.Lamins: Nuclear matrix proteins that are structural components of the NUCLEAR LAMINA. They are found in most multicellular organisms.Dimethyldithiocarbamate: A chemical that acts as a dopamine beta-hydroxylase inhibitor. Its salts are agricultural fungicides. It is inferior to diethyldithiocarbamate as a chelating agent.Nuclear Lamina: A lattice of fibrils which covers the entire inner surface of the nuclear envelope and interlinks nuclear pores (NUCLEAR PORE).Drosophila: A genus of small, two-winged flies containing approximately 900 described species. These organisms are the most extensively studied of all genera from the standpoint of genetics and cytology.Stem Cell Research: Experimentation on STEM CELLS and on the use of stem cells.Drosophila Proteins: Proteins that originate from insect species belonging to the genus DROSOPHILA. The proteins from the most intensely studied species of Drosophila, DROSOPHILA MELANOGASTER, are the subject of much interest in the area of MORPHOGENESIS and development.Research: Critical and exhaustive investigation or experimentation, having for its aim the discovery of new facts and their correct interpretation, the revision of accepted conclusions, theories, or laws in the light of newly discovered facts, or the practical application of such new or revised conclusions, theories, or laws. (Webster, 3d ed)Research Personnel: Those individuals engaged in research.Awards and PrizesResearch Support as Topic: Financial support of research activities.Biomedical Research: Research that involves the application of the natural sciences, especially biology and physiology, to medicine.Sesquiterpenes15-Oxoprostaglandin 13-Reductase: (5Z)-(15S)-11 alpha-Hydroxy-9,15-dioxoprostanoate:NAD(P)+ delta(13)-oxidoreductase. An enzyme active in prostaglandin E and F catabolism. It catalyzes the reduction of the double bond at the 13-14 position of the 15-ketoprostaglandins and uses NADPH as cofactor. EC 1.3.1.48.Fleroxacin: A broad-spectrum antimicrobial fluoroquinolone. The drug strongly inhibits the DNA-supercoiling activity of DNA GYRASE.Apoptosis: One of the mechanisms by which CELL DEATH occurs (compare with NECROSIS and AUTOPHAGOCYTOSIS). Apoptosis is the mechanism responsible for the physiological deletion of cells and appears to be intrinsically programmed. It is characterized by distinctive morphologic changes in the nucleus and cytoplasm, chromatin cleavage at regularly spaced sites, and the endonucleolytic cleavage of genomic DNA; (DNA FRAGMENTATION); at internucleosomal sites. This mode of cell death serves as a balance to mitosis in regulating the size of animal tissues and in mediating pathologic processes associated with tumor growth.Cefotaxime: Semisynthetic broad-spectrum cephalosporin.Cell Line, Tumor: A cell line derived from cultured tumor cells.Tumor Cells, Cultured: Cells grown in vitro from neoplastic tissue. If they can be established as a TUMOR CELL LINE, they can be propagated in cell culture indefinitely.

Bcl-2 regulates amplification of caspase activation by cytochrome c. (1/7010)

Caspases, a family of specific proteases, have central roles in apoptosis [1]. Caspase activation in response to diverse apoptotic stimuli involves the relocalisation of cytochrome c from mitochondria to the cytoplasm where it stimulates the proteolytic processing of caspase precursors. Cytochrome c release is controlled by members of the Bcl-2 family of apoptosis regulators [2] [3]. The anti-apoptotic members Bcl-2 and Bcl-xL may also control caspase activation independently of cytochrome c relocalisation or may inhibit a positive feedback mechanism [4] [5] [6] [7]. Here, we investigate the role of Bcl-2 family proteins in the regulation of caspase activation using a model cell-free system. We found that Bcl-2 and Bcl-xL set a threshold in the amount of cytochrome c required to activate caspases, even in soluble extracts lacking mitochondria. Addition of dATP (which stimulates the procaspase-processing factor Apaf-1 [8] [9]) overcame inhibition of caspase activation by Bcl-2, but did not prevent the control of cytochrome c release from mitochondria by Bcl-2. Cytochrome c release was accelerated by active caspase-3 and this positive feedback was negatively regulated by Bcl-2. These results provide evidence for a mechanism to amplify caspase activation that is suppressed at several distinct steps by Bcl-2, even after cytochrome c is released from mitochondria.  (+info)

C/EBPalpha regulates generation of C/EBPbeta isoforms through activation of specific proteolytic cleavage. (2/7010)

C/EBPalpha and C/EBPbeta are intronless genes that can produce several N-terminally truncated isoforms through the process of alternative translation initiation at downstream AUG codons. C/EBPbeta has been reported to produce four isoforms: full-length 38-kDa C/EBPbeta, 35-kDa LAP (liver-enriched transcriptional activator protein), 21-kDa LIP (liver-enriched transcriptional inhibitory protein), and a 14-kDa isoform. In this report, we investigated the mechanisms by which C/EBPbeta isoforms are generated in the liver and in cultured cells. Using an in vitro translation system, we found that LIP can be generated by two mechanisms: alternative translation and a novel mechanism-specific proteolytic cleavage of full-length C/EBPbeta. Studies of mice in which the C/EBPalpha gene had been deleted (C/EBPalpha-/-) showed that the regulation of C/EBPbeta proteolysis is dependent on C/EBPalpha. The induction of C/EBPalpha in cultured cells leads to induced cleavage of C/EBPbeta to generate the LIP isoform. We characterized the cleavage activity in mouse liver extracts and found that the proteolytic cleavage activity is specific to prenatal and newborn livers, is sensitive to chymostatin, and is completely abolished in C/EBPalpha-/- animals. The lack of cleavage activity in the livers of C/EBPalpha-/- mice correlates with the decreased levels of LIP in the livers of these animals. Analysis of LIP production during liver regeneration showed that, in this system, the transient induction of LIP is dependent on the third AUG codon and most likely involves translational control. We propose that there are two mechanisms by which C/EBPbeta isoforms might be generated in the liver and in cultured cells: one that is determined by translation and a second that involves C/EBPalpha-dependent, specific proteolytic cleavage of full-length C/EBPbeta. The latter mechanism implicates C/EBPalpha in the regulation of posttranslational generation of the dominant negative C/EBPbeta isoform, LIP.  (+info)

Interaction of inflammatory cells and oral microorganisms. II. Modulation of rabbit polymorphonuclear leukocyte hydrolase release by polysaccharides in response to Streptococcus mutans and Streptococcus sanguis. (3/7010)

The release of lysosomal hydrolases from polymorphonuclear leukocytes (PMNs) has been postulated in the pathogenesis of tissue injury in periodontal disease. In the present study, lysosomal enzyme release was monitored from rabbit peritoneal exudate PMNs exposed to Streptocccus mutans or Streptococcus sanguis. S. mutans grown in brain heart infusion (BHI) broth failed to promote significant PMN enzyme release. S. sanguis grown in BHI broth, although more effective than S. mutants, was a weak stimulus for promotion of PMN hydrolase release. Preincubation of washed, viable S. mutans in sucrose or in different-molecular-weight dextrans resulted in the ability of the organisms to provoke PMN release reactions. This effect could bot be demonstrated with boiled or trypsinized S. mutans or with viable S. sanguis. However, when grown in BHI broth supplemented with sucrose, but not with glucose, both S. mutans and S. sanguis triggered discharge of PMN enzymes. The mechanism(s) whereby dextran or sucrose modulates PMN-bacterial interaction may in some manner be related to promotion of microbial adhesiveness or aggregation by dextran and by bacterial synthesis of glucans from sucrose.  (+info)

Acetyl-CoA:1-O-alkyl-2-lyso-sn-glycero-3-phosphocholine acetyltransferase is directly activated by p38 kinase. (4/7010)

Acetyl-CoA:1-O-alkyl-2-lyso-sn-glycero-3-phosphocholine acetyltransferase, along with phospholipase A2, is a key regulator of platelet-activating factor biosynthesis via the remodeling pathway. We have now obtained evidence in human neutrophils indicating that this enzyme is regulated by a specific member of the mitogen-activated protein kinases, namely the p38 kinase. We earlier demonstrated that tumor necrosis factor-alpha (TNF-alpha) as well as N-formyl-methionyl-leucyl-phenylalanine treatment leads to increased phosphorylation and activation of p38 kinase in human neutrophils. Strikingly, in the present study these stimuli increased the catalytic activity of acetyltransferase up to 3-fold, whereas 4-phorbol 12-myristate 13-acetate, which activates the extracellular-regulated kinases (ERKs) but not p38 kinase, had no effect. Furthermore, a selective inhibitor of p38 kinase, SB 203580, was able to abolish the TNF-alpha- and N-formyl-methionyl-leucyl-phenylalanine-induced activation of acetyltransferase. The same effect was not observed in the presence of an inhibitor that blocked ERK activation (PD 98059). Complementing the findings in intact cells, we have shown that recombinant, activated p38 kinase added to microsomes in the presence of Mg2+ and ATP increased acetyltransferase activity to the same degree as in microsomes obtained from TNF-alpha-stimulated cells. No activation of acetyltransferase occurred upon treatment of microsomes with either recombinant, activated ERK-1 or ERK-2. Finally, the increases in acetyltransferase activity induced by TNF-alpha could be ablated by treating the microsomes with alkaline phosphatase. Thus acetyltransferase appears to be a downstream target for p38 kinase but not ERKs. These data from whole cells as well as cell-free systems fit a model wherein stimulus-induced acetyltransferase activation is mediated by a phosphorylation event catalyzed directly by p38 kinase.  (+info)

Terreic acid, a quinone epoxide inhibitor of Bruton's tyrosine kinase. (5/7010)

Bruton's tyrosine kinase (Btk) plays pivotal roles in mast cell activation as well as in B cell development. Btk mutations lead to severe impairments in proinflammatory cytokine production induced by cross-linking of high-affinity IgE receptor on mast cells. By using an in vitro assay to measure the activity that blocks the interaction between protein kinase C and the pleckstrin homology domain of Btk, terreic acid (TA) was identified and characterized in this study. This quinone epoxide specifically inhibited the enzymatic activity of Btk in mast cells and cell-free assays. TA faithfully recapitulated the phenotypic defects of btk mutant mast cells in high-affinity IgE receptor-stimulated wild-type mast cells without affecting the enzymatic activities and expressions of many other signaling molecules, including those of protein kinase C. Therefore, this study confirmed the important roles of Btk in mast cell functions and showed the usefulness of TA in probing into the functions of Btk in mast cells and other immune cell systems. Another insight obtained from this study is that the screening method used to identify TA is a useful approach to finding more efficacious Btk inhibitors.  (+info)

A multisubunit acetyl coenzyme A carboxylase from soybean. (6/7010)

A multisubunit form of acetyl coenzyme A (CoA) carboxylase (ACCase) from soybean (Glycine max) was characterized. The enzyme catalyzes the formation of malonyl CoA from acetyl CoA, a rate-limiting step in fatty acid biosynthesis. The four known components that constitute plastid ACCase are biotin carboxylase (BC), biotin carboxyl carrier protein (BCCP), and the alpha- and beta-subunits of carboxyltransferase (alpha- and beta-CT). At least three different cDNAs were isolated from germinating soybean seeds that encode BC, two that encode BCCP, and four that encode alpha-CT. Whereas BC, BCCP, and alpha-CT are products of nuclear genes, the DNA that encodes soybean beta-CT is located in chloroplasts. Translation products from cDNAs for BC, BCCP, and alpha-CT were imported into isolated pea (Pisum sativum) chloroplasts and became integrated into ACCase. Edman microsequence analysis of the subunits after import permitted the identification of the amino-terminal sequence of the mature protein after removal of the transit sequences. Antibodies specific for each of the chloroplast ACCase subunits were generated against products from the cDNAs expressed in bacteria. The antibodies permitted components of ACCase to be followed during fractionation of the chloroplast stroma. Even in the presence of 0.5 M KCl, a complex that contained BC plus BCCP emerged from Sephacryl 400 with an apparent molecular mass greater than about 800 kD. A second complex, which contained alpha- and beta-CT, was also recovered from the column, and it had an apparent molecular mass of greater than about 600 kD. By mixing the two complexes together at appropriate ratios, ACCase enzymatic activity was restored. Even higher ACCase activities were recovered by mixing complexes from pea and soybean. The results demonstrate that the active form of ACCase can be reassembled and that it could form a high-molecular-mass complex.  (+info)

Mos positively regulates Xe-Wee1 to lengthen the first mitotic cell cycle of Xenopus. (7/7010)

Several key developmental events occur in the first mitotic cell cycle of Xenopus; consequently this cycle has two gap phases and is approximately 60-75 min in length. In contrast, embryonic cycles 2-12 consist only of S and M phases and are 30 min in length. Xe-Wee1 and Mos are translated and degraded in a developmentally regulated manner. Significantly, both proteins are present in the first cell cycle. We showed previously that the expression of nondegradable Mos, during early interphase, delays the onset of M phase in the early embryonic cell cycles. Here we report that Xe-Wee1 is required for the Mos-mediated M-phase delay. We find that Xe-Wee1 tyrosine autophosphorylation positively regulates Xe-Wee1 and is only detected in the first 30 min of the first cell cycle. The level and duration of Xe-Wee1 tyrosine phosphorylation is elevated significantly when the first cell cycle is elongated with nondegradable Mos. Importantly, we show that the tyrosine phosphorylation of Xe-Wee1 is required for the Mos-mediated M-phase delay. These findings indicate that Mos positively regulates Xe-Wee1 to generate the G2 phase in the first cell cycle and establish a direct link between the MAPK signal transduction pathway and Wee1 in vertebrates.  (+info)

The retinoblastoma protein alters the phosphorylation state of polyomavirus large T antigen in murine cell extracts and inhibits polyomavirus origin DNA replication. (8/7010)

The retinoblastoma tumor suppressor protein (pRb) can associate with the transforming proteins of several DNA tumor viruses, including the large T antigen encoded by polyomavirus (Py T Ag). Although pRb function is critical for regulating progression from G1 to S phase, a role for pRb in S phase has not been demonstrated or excluded. To identify a potential effect of pRb on DNA replication, pRb protein was added to reaction mixtures containing Py T Ag, Py origin-containing DNA (Py ori-DNA), and murine FM3A cell extracts. We found that pRb strongly represses Py ori-DNA replication in vitro. Unexpectedly, however, this inhibition only partially depends on the interaction of pRb with Py T Ag, since a mutant Py T Ag (dl141) lacking the pRb interaction region was also significantly inhibited by pRb. This result suggests that pRb interferes with or alters one or more components of the murine cell replication extract. Furthermore, the ability of Py T Ag to be phosphorylated in such extracts is markedly reduced in the presence of pRb. Since cyclin-dependent kinase (CDK) phosphorylation of Py T Ag is required for its replication function, we hypothesize that pRb interferes with this phosphorylation event. Indeed, the S-phase CDK complex (cyclin A-CDK2), which phosphorylates both pRb and Py T Ag, alleviates inhibition caused by pRb. Moreover, hyperphosphorylated pRb is incapable of inhibiting replication of Py ori-DNA in vitro. We propose a new requirement for maintaining pRb phosphorylation in S phase, namely, to prevent deleterious effects on the cellular replication machinery.  (+info)

*Cell-free system

A cell-free system is an in vitro tool widely used to study biological reactions that happen within cells apart from a full ... where a cell-free translation system based on Escherichia coli (E. coli), of the cell extract-based type, had the mRNA template ... Cell-free systems may be divided into two primary classifications: cell extract-based, which remove components from within a ... Notably, in work leading to a Nobel prize the Nirenberg and Matthaei experiment used a cell-free system, of the cell extract- ...

*Prion

doi:10.1016/j.cell.2005.02.011. PMID 15851027. Karapetyan YE (Feb 2012). "Viruses do replicate in cell-free systems". ... "Stimulation of poliovirus synthesis in a HeLa cell-free in vitro translation-RNA replication system by viral protein 3CDpro". ... Studies propagating TSE infectivity in cell-free reactions and in purified component chemical reactions is thought to strongly ... However, some viruses, such as Poliovirus, have the ability to replicate in cell-free reactions. The 'virino hypothesis' ...

*History of RNA biology

Schweet R, Lamfrom H, Allen E (1958). "THE SYNTHESIS OF HEMOGLOBIN IN A CELL-FREE SYSTEM". Proc. Natl. Acad. Sci. U.S.A. 44 (10 ... LAMFROM H (1961). "Factors determining the specificity of hemoglobin synthesized in a cell-free system". J. Mol. Biol. 3: 241- ... An experimental system was developed in which an intron-containing rRNA precursor from the nucleus of the ciliated protozoan ... doi:10.1016/j.cell.2009.01.035. PMC 2675692 . PMID 19239886. Bonasio, R.; Tu, S.; Reinberg, D. (2010). "Molecular Signals of ...

*Aerobactin synthase

Appanna DL, Grundy BJ, Szczepan EW & Viswanatha T (1984). "Aerobactin synthesis in a cell-free system of Aerobacter aerogenes ...

*N2-citryl-N6-acetyl-N6-hydroxylysine synthase

Appanna, D.L.; Grundy, B.J.; Szczepan, E.W.; Viswanatha, T. (1984). "Aerobactin synthesis in a cell-free system of Aerobacter ...

*Promega

"Functional protein expression from a DNA based wheat germ cell-free system". J. Struct. Funct. Genomics. 8 (4): 199-208. doi: ... The company was an early supplier in the cell-free protein synthesis field and is continuing to develop its portfolio in this ... The company also sells its own Maxwell 16 System, a bench-top automated purification system for lower throughput research and ... The luminometers with injection systems are available for use with dual-reporter assays like the Dual-Luciferase systems. The Y ...

*Robert G. Roeder

1980: The development of cell-free systems leads to the identification of complex sets of proteins called accessory factors ... 1977-1979: Roeder develops cell-free systems to better study transcription. Composed of the purified RNA polymerases and ... the first cell-specific coactivator, discovered by Roeder in 1992, is unique to immune system B cells. 1996: Roeder's ... or specific to one particular cell type. Roeder and colleagues introduce the concept of cell specificity after they demonstrate ...

*Adenylate dimethylallyltransferase

Chen CM, Melitz DK (1979). "Cytokinin biosynthesis in a cell-free system from cytokinin-autotrophic tobacco tissue cultures". ...

*GM2A

Jinnai H, Nakamura S (2000). "Characterization of phospholipase D activation by GM2 activator in a cell-free system". Kobe ... 1]. In melanocytic cells GM2A gene expression may be regulated by MITF. Mutations in this gene, inherited in an autosomal ... GM2A is a lipid transfer protein that stimulates the enzymatic processing of gangliosides, and also T-cell activation through ... 2008). "Novel MITF targets identified using a two-step DNA microarray strategy". Pigment Cell Melanoma Res. 21 (6): 665-76. doi ...

*Extrachromosomal DNA

"Replication of Independent Formation of Extrachromosomal Circular DNA in Mammalian Cell-Free System". PLOS ONE. 4 (7): 1-8. doi ... For example, muscle and liver cells contain more copies of mtDNA per mitochondrion than blood and skin cells do. Due to the ... Chloroplasts contain multiple copies of cpDNA and the number can vary not only from species to species or cell type to cell ... For this virus to persist, the circular genome must be replicated and inherited during cell division. Cells can recognize ...

*Aryl-alcohol dehydrogenase (NADP+)

Reduction of aromatic acids to aldehydes and alcohols in the cell-free system. 2. Purification and properties of aryl-alcohol: ...

*GPLD1

Jinnai H, Nakamura S (2000). "Characterization of phospholipase D activation by GM2 activator in a cell-free system". Kobe ... Cell Biol. 80 (2): 253-60. doi:10.1139/o02-004. PMID 11989719. Xiaotong H, Hannocks MJ, Hampson I, Brunner G (2002). "GPI- ... The GPI-anchor is a glycolipid found on many blood cells. The protein encoded by this gene is a GPI degrading enzyme. ... 1994). "Release of GPI-anchored membrane proteins by a cell-associated GPI-specific phospholipase D". EMBO J. 13 (7): 1741-51. ...

*Aryl-aldehyde dehydrogenase (NADP+)

Gross GG, Zenk MH (1969). "[Reduction of aromatic acids to aldehydes and alcohols in the cell-free system. 1. Purification and ...

*CLSPN

Clarke CA, Clarke PR (2005). "DNA-dependent phosphorylation of Chk1 and Claspin in a human cell-free system". Biochem. J. 388 ( ... Cell. 23 (3): 319-29. doi:10.1016/j.molcel.2006.06.013. PMID 16885022. Mamely I, van Vugt MA, Smits VA, et al. (2006). "Polo- ... Cell. Biol. 26 (16): 6056-64. doi:10.1128/MCB.00492-06. PMC 1592810 . PMID 16880517. Mailand N, Bekker-Jensen S, Bartek J, ... Cell. 23 (3): 307-18. doi:10.1016/j.molcel.2006.06.016. PMID 16885021. Peschiaroli A, Dorrello NV, Guardavaccaro D, et al. ( ...

*Cell-free protein synthesis

... is the production of protein using biological machinery in a cell-free system, that is, without the use of living cells. The in ... "High-Level Cell-Free Production of Membrane Proteins with Nanodiscs". Cell-Free Protein Synthesis. Methods in Molecular Biology ... They used a cell-free system to translate a poly-uracil RNA sequence (or UUUUU... in biochemical terms) and discovered that the ... Spirin, A.; Baranov, V.; Ryabova, L.; Ovodov, S.; Alakhov, Y. (1988). "A continuous cell-free translation system capable of ...

*Genetic code

They used a cell-free system to translate a poly-uracil RNA sequence (i.e., UUUUU...) and discovered that the polypeptide that ... ISBN 0-465-09138-5. Nirenberg MW, Matthaei JH (Oct 1961). "The dependence of cell-free protein synthesis in E. coli upon ... It was a (single cell) bacterium with two synthetic bases (called X and Y). The bases survived cell division. In 2017 a mouse ... Biology of the Cell / Under the Auspices of the European Cell Biology Organization. 95 (3-4): 169-78. doi:10.1016/S0248-4900(03 ...

*Nirenberg and Matthaei experiment

In order to decipher this biological mystery, Nirenberg and Matthaei needed a cell-free system that would build amino acids ... The experimentation with synthetic RNA in a cell-free system was a key technical innovation. In 1961, when they announced their ... Nirenberg, M.W. & Matthaei, H.J. (1961). "The Dependence Of Cell- Free Protein Synthesis In E. coli Upon Naturally Occurring Or ... In the experiment, an extract from bacterial cells that could make protein even when no intact living cells were present was ...

*RNA-induced silencing complex

"Let-7 microRNA-mediated mRNA deadenylation and translational repression in a mammalian cell-free system". Genes & Development. ... S2 cells were then transfected with Drosophila cyclin E dsRNA. Cycline E is an essential gene for cell cycle progression into ... Bartel DP (2009). "MicroRNAs: target recognition and regulatory functions". Cell. 136 (2): 215-233. doi:10.1016/j.cell.2009.01. ... as suggested by data from other systems), Drosophila S2 cells were transfected with either Drosophila cyclin E dsRNAs or lacZ ...

*Let-7 microRNA precursor

"Let-7 microRNA-mediated mRNA deadenylation and translational repression in a mammalian cell-free system". Genes Dev. 21 (15): ... Dröge P, Davey CA (2008). "Do cells let-7 determine stemness?". Cell Stem Cell. 2 (1): 8-9. doi:10.1016/j.stem.2007.12.003. ... "let-7 regulates self renewal and tumorigenicity of breast cancer cells". Cell. 131 (6): 1109-23. doi:10.1016/j.cell.2007.10.054 ... and IL6 links inflammation to cell transformation". Cell. 139 (4): 693-706. doi:10.1016/j.cell.2009.10.014. PMC 2783826 . PMID ...

*L-lysine 6-monooxygenase (NADPH)

Goh CJ, Szczepan EW, Menhart N, Viswanatha T (1989). "Studies on lysine:N6-hydroxylation by cell-free systems of Aerobacter ...

*Methylobacterium organophilum

"Hemisynthesis of deuteriated adenosylhopane and conversion into bacteriohopanetetrol by a cell-free system from ...

*Nuclear pore

"Steps in the assembly of replication-competent nuclei in a cell-free system from Xenopus eggs". The Journal of Cell Biology. ... Interphase cells must also keep up a level of NPC generation to keep the levels of NPC in the cell constant as some may get ... For example, cycling mammalian and yeast cells double the amount of NPC in the nucleus between the G1 and G2 phase of the cell ... Some cells can even increase the NPC numbers due to increased transcriptional demand. There are several theories as to how NPCs ...

*Super-resolution microscopy

"Coupling between clathrin-dependent endocytic budding and F-BAR-dependent tubulation in a cell-free system". Nature Cell ... Images of cell nuclei and mitotic stages recorded with 3D-SIM. Comparison confocal microscopy - 3D-SIM Cell nucleus in prophase ... Optical resolution of cellular structures in the range of about 50 nm can be achieved even in label-free cells using the ... Up to now, the best quality in a 4Pi microscope was reached in conjunction with the STED principle in fixed cells and RESOLFT ...

*Selective progesterone receptor modulator

"Analysis of the mechanism of steroid hormone receptor-dependent gene activation in cell-free systems". Endocrine Reviews. 13 (3 ... Finally, the overall ratio of concentrations of coactivator to corepressor may differ in different cell types. At the turn of ... However, when an antagonist, e.g. mifepristone, interacts with this hydrogen bond system then its dimethylamine group clashes ... McKenna NJ, O'Malley BW (Feb 2002). "Combinatorial control of gene expression by nuclear receptors and coregulators". Cell. 108 ...

*Luis Federico Leloir

... he produced an active cell-free system, a first in scientific research. It had initially been assumed that in order to study a ... I discovered (no, not me: my team) the function of sugar nucleotides in cell metabolism. I want others to understand this, but ... Along the way, Muñoz and Leloir, unable to procure the costly centrifuge needed to separate cell contents, improvised by ... cell, scientists could not separate it from its host organism, as oxidation could only occur in intact cells. ...

*Ariosa v. Sequenom

6,258,540, which claims methods of using cell-free fetal DNA (cffDNA) circulating in maternal plasma (cell-free blood) to ... 101 progressively weakens the patent system, especially in the life sciences. And each case seems to present a new low in terms ... where development of useful new diagnostic and therapeutic methods is driven by investigation of complex biological systems. I ... he panel's decision striking down Sequenom's noninvasive prenatal test strikes at the very heart of the patent system. ...
TY - JOUR. T1 - Hemoglobin transition in erythrocytes of developing chick. Studies with cell-free protein-synthesizing systems. AU - Henderson, A. Burl. AU - Lee, John C.. PY - 1976. Y1 - 1976. N2 - Cell-free hemoglobin-synthesizing systems from erythrocytes of 4- and 17-day chick embryos have been developed. These systems have been used to investigate possible structural and functional differences in factors involved in protein synthesis obtained from these different developmental stages. Each cell-free system consists of three major cellular fractions i.e., the S-100 supernatant, the salt-washed ribosomes, and the 0.5 m KCl ribosomal wash. When the ribosomal wash fraction from one developmental stage is included in a cell-free system containing ribosomes and S-100 supernatant from the other developmental stage, a drastic reduction in the kinetics of [3H]leucine incorporation into globin products is observed, when compared to the homologous control cell-free systems. A similar depression of the ...
Principal Investigator:SUYAMA Akira, Project Period (FY):2011-04-01 - 2016-03-31, Research Category:Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Project Area:Synthetic biology for the comprehension of biomolecular networks
TY - JOUR. T1 - Characterization of cell free synthesis of collagen by lung polysomes in a heterologous system. AU - Collins, J. F.. AU - Crystal, Ronald. PY - 1975/12/1. Y1 - 1975/12/1. N2 - In normal lung growth, post pneumonectomy lung growth, and in possibly several lung disorders, there are marked alterations in the density of collagen and changes in the rate of synthesis of collagen relative to the synthesis of other lung proteins. To provide a technology to begin to understand these changes at the molecular level, polysomes were prepared from rabbit lung and translated in a heterologous cell free system including rabbit reticulocyte 0.5 m KCl ribosomal wash fraction and liver tRNA. Collagen was shown in the cell free product by collagenase sensitivity, hydroxylation of incorporated proline by peptidyl prolyl hydroxylase, agarose gel chromatography, and sodium dodecyl sulfate acrylamide gel electrophoresis. The cell free system was optimized with respect to K+, Mg2+, amino acids, and ...
Video articles in JoVE about escherichia coli include The Multifaceted Benefits of Protein Co-expression in Escherichia coli, Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology, Quantification of the Abundance and Charging Levels of Transfer RNAs in Escherichia coli, Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach, Mapping Bacterial Functional Networks and Pathways in Escherichia Coli using Synthetic Genetic Arrays, Non-Invasive Model of Neuropathogenic Escherichia coli Infection in the Neonatal Rat, Method for Labeling Transcripts in Individual Escherichia coli Cells for Single-molecule Fluorescence In Situ Hybridization Experiments, Residue-specific Incorporation of Noncanonical Amino Acids into Model Proteins Using an Escherichia coli Cell-free Transcription-translation System, Detection of Live Escherichia coli O157:H7 Cells by PMA-qPCR,
Hear from James about promising career paths in cell-free systems, and get a preview of his upcoming talk at the Cell Free Systems Conference.
Engineered antibodies are key players in therapy, diagnostics and research. In addition to full size immunoglobulin gamma (IgG) molecules, smaller formats of recombinant antibodies, such as single-chain variable fragments (scFv) and antigen binding fragments (Fab), have emerged as promising alternatives since they possess different advantageous properties. Cell-based production technologies of antibodies and antibody fragments are well-established, allowing researchers to design and manufacture highly specific molecular recognition tools. However, as these technologies are accompanied by the drawbacks of being rather time-consuming and cost-intensive, efficient and powerful cell-free protein synthesis systems have been developed over the last decade as alternatives. So far, prokaryotic cell-free systems have been the focus of interest. Recently, eukaryotic in vitro translation systems have enriched the antibody production pipeline, as these systems are able to mimic the natural pathway of antibody
The primary objective of our work was to find a practical solution to the limitations of GPCR expression imposed by heterologous systems. Although relatively large amounts of membrane proteins can be potentially produced in cellular systems, usually a small proportion becomes associated to the membrane [22]. Particularly, GPCRs are confronted to a complex array of trafficking signals, post-translational modifications, and transport systems before reaching the final destination, the plasma membrane. In addition, differences in the lipid bilayer composition and maximal tolerated membrane protein loads can additionally affect the correct insertion, folding, and yield of recombinant GPCRs.. In order to overcome these difficulties, we developed a cell-free expression system supplemented with planar membranes [15]. Although, the approach excels in expressing soluble membrane protein products, it fails to produce functional GPCRs. We favor the absence of a functional translocon machinery embedded in ...
Malaria is a major public health burden and developing the next generation of anti-malarials is vital to control the spread of disease. Protein arrays, which can investigate binding of many proteins to various probes simultaneously, are becoming an important tool in the drug development process. Results from protein arrays can be affected by the sample purity, as high amounts of protein from the translation system can mask positive interactions on the arrays. The results would be improved by separating the malarial protein from the translation system proteins. However, purifying the hundreds of proteins for arrays using traditional affinity fusion tags is extremely time-consuming. To reduce the time required to produce protein samples at the desired purity, enrichment schemes were developed that covalently attach a small molecule to the proteins in the wheat cell-free expression system. After translation, separation matrices with high specificity for the small molecule modification were added to ...
Kits and reagents for cell-free protein expression in just a few hours using mRNA templates in translational systems, or DNA template (plasmid DNA or PCR fragments) in coupled transcription and translation systems.
Creative Biostructure offers advanced custom Mempro™ CoA-Dependent acyltransferases production services using cell-free expression system.
The M species (medium sized) dsRNA (1.1-1.4 x 10(6) daltons) isolated from a toxin-producing yeast killer strain (K+R+) and three related, defective interfering (suppressive) S species dsRNAs of the yeast killer-associated cytoplasmic multicomponent viral-like particle system were analyzed by in vitro translation in a wheat germ cell-free protein synthesis system. Heat-denatured M species dsRNA programmed the synthesis of two major polypeptides, M-P1 (32,000 daltons) and M-P2 (30,000 daltons). M-P1 has been shown by the criteria of proteolytic peptide mapping and cross-antigenicity to contain ihe 12,000 dalton polypeptide corresponding to the in vivo produced killer toxin, thus establishing thiat it is the M species dsRNA which carries the toxin gene. An M species dsRNA obtained from a neutral strain (K-R+) also programmed the in vitro synthesis of a polypeptide identical in molecular weight to M-P1, thus indicating that the cytoplasmic determinant of the mutant neutral phenotype is either a simple
Cell-free protein synthesis is a powerful method to explore the structure and function of membrane proteins and to analyze the targeting and translocation of proteins across the ER membrane. Developing a cell-free system based on cultured cells for the synthesis of membrane proteins could provide a highly reproducible alternative to the use of tissues from living animals. We isolated Sf21 microsomes from cultured insect cells by a simplified isolation procedure and evaluated the performance of the translocation system in combination with a cell-free translation system originating from the same source. The isolated microsomes contained the basic translocation machinery for polytopic membrane proteins including SRP-dependent targeting components, translocation channel (translocon)-dependent translocation, and the apparatus for signal peptide cleavage and N-linked glycosylation. A transporter protein synthesized with the cell-free system could be functionally reconstituted into a lipid bilayer. In addition
Quantitative proteomic approaches using selected reaction monitoring (SRM) are currently limited by the difficulty in the preparation of reference standards. In this study, we demonstrat the high-throughput production of a reference peptide library using a wheat germ cell-free synthesis system to develop a l 2016 Hot Articles in Molecular BioSystems
[Problem] To provide a method for detecting a myriad of proteins that contribute to autoimmune diseases with high sensitivity and high efficiency, and a method for analyzing the data obtained as a result of the detection method. [Solution] In order to construct the abovementioned detection method and analysis method, a means is provided for detecting autoantibody production by bringing mammal-derived proteins derived, which are expressed by a cell-free protein synthesis system, into contact with samples derived from patients with autoimmune diseases, and for comprehensively analyzing the proteins that contribute to autoimmune diseases by statistically analyzing the detected data, and performing gene ontology analysis and/or pathway analysis.
... ,System Fibonacci FX 1.0.1.0 is Is the advanced deluxe version of the basic System Fibonacci software.
Do you find that your PC is acting weirdly lately with frequent message popping up? smschk.exe might be the culprit here. PC is a complicated machine and with so many different files, settings and procedures to monitor, it is hard to identify just what is slowing you down not to mention implementing the correct technical changes to recover the loss in performance. Firstly, its very important to identify the error that is causing the slow down and lacklustre performance.. Recommendation for Would it be smschk.exe or other hidden PC errors that is playing prank? Find out here with the FREE system scan. From our experience, smschk.exe is most likely a virus or trojan. It is highly recommended that you run a FREE system scan to automatically optimize your registry, memory CPU and your PC settings. Damage to your computers registry could be compromising your PCs performance and causing system breakout and crashes. Dont risk it! We recommend that you run this FREE system scan to identify ...
Do you find that your PC is acting weirdly lately with frequent message popping up? direct3d.exe might be the culprit here. PC is a complicated machine and with so many different files, settings and procedures to monitor, it is hard to identify just what is slowing you down not to mention implementing the correct technical changes to recover the loss in performance. Firstly, its very important to identify the error that is causing the slow down and lacklustre performance.. Recommendation for Would it be direct3d.exe or other hidden PC errors that is playing prank? Find out here with the FREE system scan. From our experience, direct3d.exe is most likely a virus or trojan. It is highly recommended that you run a FREE system scan to automatically optimize your registry, memory CPU and your PC settings. Damage to your computers registry could be compromising your PCs performance and causing system breakout and crashes. Dont risk it! We recommend that you run this FREE system scan to identify ...
Genomics-based target identification and screening using cell free systems has been the dominating principle in cancer drug discovery during the recent decade [6]. As an alternative to this approach the use of phenotype-cell-based screening may provide some distinct advantages [35]. We here performed a conditional screen with the aim of identifying compounds that are cytotoxic to multidrug resistant myeloma cells. A chemically diverse compound library was used for this purpose. The screening hit RH02104/VLX40 was the only compound that fulfilled the pre-determined criteria of a SI less than 50% in myeloma 8226/Dox40 and more than 50% in parental RPMI 8226 cells. In validation experiments VLX40 was found the difference was, albeit statistically significant, small. It can not be excluded that subtle differences in drug uptake and proliferation characteristics of the cell lines, not related to drug transporters, could contribute to the difference observed.. For exploration of mechanisms of action ...
This lecture will focus on cell free DNA in lung cancer. Topics covered will include the origin and biology of cell free DNA in this disease. Analytic and clinical validity studies, as well as evidence of clinical utility in medical decision making, will also be discussed.
Review: iDisksoft Card Recovery is powerful and professional data retrieval software tool, which can help you to recover lost or deleted photo, video and other media files easily from Memory Card: SD Card, CF Card, Digital Camera, etc..
Assay of cell-free DNA in blood offers an approach to assessment of tumor DNA. We sought to determine whether Epstein-Barr virus (EBV) DNA in cell-free blood is also a good surrogate for the presence of tumor DNA in children with Hodgkin lymphoma, as it is in adults, and whether it correlates with pediatric outcomes. Pediatric patients enrolled in a Childrens Oncology Group trial (AHOD0031) were studied at baseline and at 8 days after the initiation of treatment. At baseline, EBV DNA in cell-free blood correlated with the presence of EBV in tumor, and EBV DNA 8 days after the initiation of therapy predicted inferior event-free survival ...
Since the discovery of cell-free DNA (cfDNA) in human blood, most studies have focused on diagnostic and prognostic uses of these markers for solid tumors. Except for some prenatal tests and BEAMing, cfDNA analysis has not ...
Cancer researchers have an exciting new tool at their disposal: circulating cell-free DNA (ccfDNA) collected in minimally invasive liquid biopsies. With the potential to provide real-time mutational information about primary and metastatic tumors, cfDNA has significant potential for the detection and monitoring of biomarkers for cancer and other diseases. ...
Cell-free protein synthesis methods have been developed for production of homogeneous therapeutic proteins, including Antibody Drug Conjugates (ADCs). Many variants can be expressed in hours and rapidly assessed for function. Within days, production of chosen variants can be scaled using the same platform to generate material for clinical studies. The power and utility of the platform to design and manufacture single species antigen-targeted combination warheads will be described. In particular, the stability and pharmacokinetics of these homogeneous ADCs will also be highlighted. ...
Qualitative and quantitative testing of circulating cell free DNA (CCFDNA) can be applied for the management of malignant and benign neoplasms. Detecting circulating DNA in cancer patients may help develop a DNA profile for early stage diagnosis in malignancies. The technical issues of obtaining, using, and analyzing CCFDNA from blood will be discussed.
Principal Investigator:SHIODA Masaki, Project Period (FY):1995 - 1997, Research Category:Grant-in-Aid for Scientific Research (C), Section:一般, Research Field:Cell biology
본 kit는 반응튜브 내에서 template DNA를 이용하여 3시간 만에 원하는 단백질의 발현을 확인할 수 있는 kit로서 E. coli extract와 Master mix를 포함하고 있습니다. 이 중 E. coli extract는 T7 RNA polymerase, ribosome등이 포함되어 있고, Master mix에는 아미노산, NTPs, 에너지원등이 포함되어 template DNA만 준비되어 있으면 손쉽게 목적 단백질의 발현을 확인할 수 있습니다.
The influence of inoculation dilution and cell-free culture fluids from Vp cultures on Vp growth rates.Cultures were grown in pre-warmed MB at 37 °C. Represe
Alectinib (CH5424802) is a potent ALK inhibitor with IC50 of 1.9 nM in cell-free assays, sensitive to L1196M mutation and higher selectivity for ALK than PF-02341066, NVP-TAE684 and PHA-E429.
Nintedanib (BIBF 1120) is a potent triple angiokinase inhibitor for VEGFR1/2/3, FGFR1/2/3 and PDGFRα/β with IC50 of 34 nM/13 nM/13 nM, 69 nM/37 nM/108 nM and 59 nM/65 nM in cell-free assays. Phase 3.
Orchestrated membrane remodeling creates flows of substance and information to sustain the lifecycle of a cell. Therefore understanding the molecular mechanisms...
Reproducible experiments owing to: a defined 500 µm cell free gap, no leakage during cultivation, and no material being left behind after the insert s ...
Vincent Noireaux got his B.Sc. in applied physics at the University of Tours (France) in 1994. In 1995 he moved to Paris for graduate school at the University Paris 11 (Orsay), in physics. He did his PhD at the Curie Institute (Paris, 1996-2000) in the laboratory of Jacques Prost on the motion of the bacterium Listeria. He studied the actin cytoskeleton mechanism involved in cell motility. He learned the biology related to this project in the laboratory of Daniel Louvard. In 2000 he joined the laboratory of Albert Libchaber at the Rockefeller University in New York City where he spent five years as a postdoc. He used cell-free expression systems to construct elementary gene networks and artificial cell systems. In 2005, he moved to the University of Minnesota where he is pursuing his work in synthetic biology using cell-free expression to construct and to characterize complex biochemical systems in vitro ...
TY - JOUR. T1 - Advances in cell-free protein array methods. AU - Yu, Xiaobo. AU - Petritis, Brianne. AU - Duan, Hu. AU - Xu, Danke. AU - LaBaer, Joshua. PY - 2018/1/2. Y1 - 2018/1/2. N2 - Introduction: Cell-free protein microarrays represent a special form of protein microarray which display proteins made fresh at the time of the experiment, avoiding storage and denaturation. They have been used increasingly in basic and translational research over the past decade to study protein-protein interactions, the pathogen-host relationship, post-translational modifications, and antibody biomarkers of different human diseases. Their role in the first blood-based diagnostic test for early stage breast cancer highlights their value in managing human health. Cell-free protein microarrays will continue to evolve to become widespread tools for research and clinical management. Areas covered: We review the advantages and disadvantages of different cell-free protein arrays, with an emphasis on the methods ...
Autori: Bogdan Bancia Editorial: 2009.. Rezumat:. The three-dimensional structure determination of proteins represents an important step towards understanding their biological function and thus their roles in living organisms. Using a combination of multidimensional NMR techniques three different biomolecules were analyzed in the present study, E. coli peptidyl - prolyl cis-trans isomerase PpiB, proinsulin connecting peptide and DnaG-C. 15N-HSQC spectra were recorded of PpiB which had been expressed without further purification in a cell-free expression system with amino acid selective isotope labelling. Comparison of spectra before and after ultrafiltration indicated that labelled metabolic by-products are of low molecular weight. Therefore, the labelled protein signals are easily distinguished from those of metabolites. The structure analysis of the proinsulin connecting peptide included the assignment of 1H, 13C and 15N NMR resonances using 2D NMR, measurement of T1 (1H) relaxation times and ...
Cytokinesis, when two daughter cells are physically separated from one another, is the final stage of cell division. How dividing cells signal where the cleavage furrow should be during cytokinesis has long interested cell biologists. A major stumbling block to probing the underlying mechanisms has been the lack of a cell-free and fully controllable experimental system.. In a new paper appearing in Science (10 October 2014, Science 346 (6206):244-247), Phuong Nguyen and Aaron Groen, along with their colleagues at Harvard Medical School (Boston, MA), have reconstituted the organization of cytokinesis signaling outside living cells, using a the cell-free Xenopus system. The authors examined the biophysics involved in spatial signaling during cytokinesis using powerful imaging techniques, taking advantage of microtubule cytokinesis zones that they assembled on the surface of a microscope slide. These cytokinesis zones are relatively large (~20 mm) in the large Xenopus egg (c. 10x larger than ...
Norovirus vaccine development largely depends on recombinant virus-like-particles (VLPs). Norovirus VLPs have been produced in several cell-based expression systems with long production times. Here we report, for the first time, that norovirus VLPs can be expressed and assembled by using a cell-free protein expression system within four hours ...
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During the STTR phase I and phase II NSF projects, FAB reported that certain proteins could be made in a soluble form through a cell-free system that was controllable and efficient. With the current award, FAB will continue to develop the technology with a market-driven, therapeutic protein as FABs first target product. ...
The T7 bacteriophage RNA polymerase (T7 RNAP) serves as a model for understanding RNA synthesis, as a tool for protein expression, and as an actuator for synthetic gene circuit design in bacterial cells and cell-free extract. T7 RNAP is an attractive tool for orthogonal protein expression in bacteria owing to its compact single subunit structure and orthogonal promoter specificity. Understanding the mechanisms underlying T7 RNAP regulation is important to the design of engineered T7-based transcription factors, which can be used in gene circuit design. To explore regulatory mechanisms for T7 RNAP-driven expression, we developed a rapid and cost-effective method to characterize engineered T7-based transcription factors using cell-free protein synthesis and an acoustic liquid handler. Using this method, we investigated the effects of the tetracycline operators proximity to the T7 promoter on the regulation of T7 RNAP-driven expression. Our results reveal a mechanism for regulation that functions ...
The 5´terminal m7G cap present on most eukaryotic mRNAs promotes translation in vitro at the initiation level. For most RNAs, elimination of the cap structure causes a loss of stability, especially against exonuclease degradation, and a decrease in the formation of the initiation complex of mRNAs for protein synthesis. Certain prokaryotic mRNAs containing a 5´ terminal cap structure are translated as efficiently as or more efficiently than eukaryotic mRNAs in a eukaryotic cell-free protein synthesizing system. Also a cap requirement has been observed for splicing eukaryotic substrate RNAs. A method for efficient in vitro synthesis of capped RNA using E. coli RNA polymerase primed with m7G(5´ )ppp(5´ )G or m7G(5´ )ppp(5´ )A has been developed by Contreas et al.
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Thank you for your interest in spreading the word about Biochemical Journal.. NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.. ...
Cell-free DNA is detected in blood in many diseases, but also in healthy individuals. Cell-free DNA can originate from necrotic cells or apoptotic …
cell free DNA and maintaining stability - posted in Molecular Biology: Hey, Im looking to start examining cell free DNA and Im worried about its purported short half life. Can anyone give me a working estimate on its half life/stability?? Has anyone used reagents like Invitrogens RNALater or Qiagens Allprotect tissue reagent, to maintain the integrity of cfDNA for longer periods of time? thanks!!
This report focuses on the global Cell Free Protein Expression status, future forecast, growth opportunity, key market and key players. The study objectives are to present the Cell Free Protein Expression development in United States, Europe and China.
STRO-001 was developed with Sutros proprietary cell-free protein synthesis and site-specific conjugation platforms, which facilitate multiple rounds of antibody and ADC optimization," said Dr. Arturo Molina, a medical oncologist and Sutros chief medical officer.. "Sutros Xpress CF+™ platform enables it to produce novel ADCs that directly target cancer cells and avoid a toxic bystander effect on adjacent healthy cells," he added.. Unlike conventional cell-based expression systems, Sutros technology isolates a cells protein production machinery into a cell-free extract, Xtract CF™, which includes all the necessary biochemical components for energy production, transcription and translation. The Xpress CF+™ platform further supports incorporation of non-natural amino acids in specific positions in the protein of interest, allowing for site-specific conjugation of cytotoxins and the creation of homogeneous ADCs. This process is capable of producing single proteins at large scale within ...
A rapid method for gene expression analysis, PURExpress® is a novel cell-free transcription/translation system reconstituted from the purified components necessary for E. coli translation. The relative nuclease-free and protease-free nature of the PURExpress platform preserves the integrity of DNA and RNA templates/ complexes and results in proteins that are free of modification and degradation. Transcription and translation are carried out in a one-step reaction, and require the mixing of only two tubes. With results available in a few hours, PURExpress saves valuable laboratory time and is ideal for high throughput technologies.
The purpose was to investigate total cell-free DNA (cfDNA) in colorectal cancer (CRC) patients during treatment with second-line chemotherapy and in healthy controls and patients with different comorbidities. Patient treated with second-line irinotecan for metastatic CRC (n = 100), a cohort of healthy controls with and without comorbidity (n = 70 and 100, respectively) were included. cfDNA was quantified by an in-house developed quantitative polymerase chain reaction from plasma samples drawn prior to the first cycle of chemotherapy and at time of progression. cfDNA levels were significantly higher in CRC compared to controls, with a clear capability for discriminating between the groups (receiver operation curve analysis; area under the curve 0.82, p , 0.0001). Patients with high levels had a shorter survival from irinotecan compared to those with lover levels. The cohort independent upper normal limit divided patients into high and low risk groups. The progression-free survival (PFS) was 2.1 ...
1) 1st PCR: The DNA encoding a protein of interest will be amplified. The PCR product will contain start and stop codon, whole coding region of the protein, and additional sequences for the 2nd PCR. The primers for this step are gene specific primers with overhangs and can be ordered on the Bioneers website.. 2) 2nd PCR: Using a product from the 1st PCR, 2nd PCR performed using primer set by which T7 promoter and RBS are introduced at its 5 end and T7 terminator is added at its 3 end. PCR product after 2nd PCR should be gel-purified for the cell-free protein synthesis.. ...
Cyanine fluorophores are encoded as non-canonical amino acids to produce functional proteins in cell-free translation systems and live cells for single-molecule imaging.
Erved at an apparent molecular mass higher than the predicted 127 kDa in COS-7, HeLa, 293 Cells and in cell free transcription/translation systems (J. Perdomo,
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Zu diesem lizenzpflichtigen Artikel gibt es eine Open Access Version, die kostenlos und ohne Lizenzbeschränkung gelesen werden kann. Die Open Access Version kann inhaltlich von der lizenzpflichtigen Version abweichen ...
Schematic showing the infection process. The viral transmissions used cell-free supernatants. The designations K1 through K7 dont represent the order of the ex
Product Description:. SGI-1776 free base is a novel ATP competitive inhibitor of Pim1 with IC50 of 7 nM in a cell-free assay, 50- and 10-fold selective versus Pim2 and Pim3, also potent to Flt3 and haspin. Phase 1. ...
Video articles in JoVE about bacteriophage t7 include Kinetics of Lagging-strand DNA Synthesis In Vitro by the Bacteriophage T7 Replication Proteins, Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology, Synthesis of Infectious Bacteriophages in an E. coli-based Cell-free Expression System, Rescue of Recombinant Newcastle Disease Virus from cDNA, Expression, Isolation, and Purification of Soluble and Insoluble Biotinylated Proteins for Nerve Tissue Regeneration, Reverse Genetics Mediated Recovery of Infectious Murine Norovirus, Whole Mount RNA Fluorescent in situ Hybridization of Drosophila Embryos, Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay, Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins, Using Tomoauto: A Protocol for High-throughput Automated Cryo-electron Tomography, Visualizing Single-molecule
AB - The polyhydroxyalkanoates (PHAs) are microbially-produced biopolymers that could potentially be used as sustainable alternatives to oil-derived plastics. However, PHAs are currently more expensive to produce than oil-derived plastics. Therefore, more efficient production processes would be desirable. Cell-free metabolic engineering strategies have already been used to optimise several biosynthetic pathways and we envisioned that cell-free strategies could be used for optimising PHAs biosynthetic pathways. To this end, we developed several Escherichia coli cell-free systems for in vitro prototyping PHAs biosynthetic operons, and also for screening relevant metabolite recycling enzymes. Furthermore, we customised our cell-free reactions through the addition of whey permeate, an industrial waste that has been previously used to optimise in vivo PHAs production. We found that the inclusion of an optimal concentration of whey permeate enhanced relative cell-free GFPmut3b production by ∼50%. In ...
A living cell has numerous proteins, only a few thousand of which have been identified to date. Cell-free protein synthesis is a useful and promising technique to discover and produce various proteins that might be beneficial for biotechnological, pharmaceutical, and medical applications. For this study, we evaluated the performance and the general applicability of our previously developed microreactor array chip to cell-free protein synthesis by comparisons with a commercially available system. The microreactor array chip comprises a temperature control chip made of glass and a disposable reaction chamber chip made of polydimethylsiloxane (PDMS). For evaluation of the microreactor array chip, rat adipose-type fatty acid binding protein, glyceraldehyde-3-phosphate dehydrogenase, cyclophilin, and firefly luciferase were synthesized from their respective DNA templates using a cell-free extract prepared from ,i,Escherichia coli,/i,. All these proteins were synthesized in the microreactor array ...
Engineered gene circuits offer an opportunity to harness biological systems for biotechnological and biomedical applications. However, reliance on native host promoters for the construction of circuit elements, such as logic gates, can make the implementation of predictable, independently functioning circuits difficult. In contrast, T7 promoters offer a simple orthogonal expression system for use in a variety of cellular backgrounds and even in cell-free systems. Here we develop a T7 promoter system that can be regulated by two different transcriptional repressors for the construction of a logic gate that functions in cells and in cell-free systems. We first present LacI repressible T7lacO promoters that are regulated from a distal lac operator site for repression. We next explore the positioning of a tet operator site within the T7lacO framework to create T7 promoters that respond to tet and lac repressors and realize an IMPLIES gate. Finally, we demonstrate that these dual input sensitive promoters
Limitations on biofuel production using cell culture (Escherichia coli, Clostridium, Saccharomyces cerevisiae, brown microalgae, blue-green algae and others) include low product (alcohol) concentrations (≤0.2 vol%) due to feedback inhibition, instability of cells, and lack of economical product recovery processes. To overcome these challenges, an alternate simplified biofuel production scheme was tested based on a cell-free immobilized enzyme system. Using this cell free system, we were able to obtain about 2.6 times higher concentrations of iso-butanol using our non-optimized system as compared with live cell systems. This process involved two steps: (i) converts acid to aldehyde using keto-acid decarboxylase (KdcA), and (ii) produces alcohol from aldehyde using alcohol dehydrogenase (ADH) with a cofactor (NADH) conversion from inexpensive formate using a third enzyme, formate dehydrogenase (FDH). To increase stability and conversion efficiency with easy separations, the first two enzymes ...
Sigma-Aldrich offers abstracts and full-text articles by [Christy Catherine, Seung-Won Lee, Jung Won Ju, Ho-Cheol Kim, Hyun-Il Shin, Yu Jung Kim, Dong-Myung Kim].
We propose a probabilistic method, CancerLocator, which exploits the diagnostic potential of cell-free DNA by determining not only the presence but also the location of tumors. CancerLocator simultaneously infers the proportions and the tissue-of-origin of tumor-derived cell-free DNA in a blood sample using genome-wide DNA methylation data. CancerLocator outperforms two established multi-class classification methods on simulations and real data, even with the low proportion of tumor-derived DNA in the cell-free DNA scenarios. CancerLocator also achieves promising results on patient plasma samples with low DNA methylation sequencing coverage.
KINGS COLLEGE, LONDON Charities. Drs N. Abbott and G. Connolly and Prof P. McNaughton, Pounds 43,005 from the Wellcome Trust (signalling between cells associated with the cerebral vasculature: the role of purines and pyrimidines); Dr G. Jones, Pounds 108,365 from the Arthritis and Rheumatism Council (role of Rac and Rho proteins in the chemotaxis-driven migration of macrophages); Dr S. Persaud, Pounds 92,705 from the Wellcome Trust (identification by insertional mutagenesis of genes involved in the regulation of pancreatic beta cells); Dr R. Brooks, Pounds 1,313 from the Wellcome Trust (initiation of DNA synthesis in vertebrate cells: development of cell free system); Dr S. Wilson, Pounds 540,755 from the Wellcome Trust (fellowship in basic biomedical science science - analysis of mechanisms underlying regional patterning of the embryonic vertebrate forebrain); Prof V. Preedy, Pounds 54,491 from the Wellcome Trust (a study of transcriptional control of protein synthesis in alcohol-induced ...
633 Best Hydrogen Fuel Cell Free Vector Art Downloads from the Vecteezy community. Hydrogen Fuel Cell Free Vector Art licensed under creative commons, open source, and more!
Genomic DNA sequences in cell-free plasma are biomarkers of cancer prognosis, where characteristic changes in methylation of tumour suppressor or oncogene DNA regions are indicative of changes in gene activity. Also, cell-free fetal DNA can be distinguished, by its methylation status, from the maternal DNA in the plasma of pregnant women, hence providing DNA biomarkers for the proposed minimally-invasive diagnosis of fetal aneuploidies, including Downs syndrome. However, the production and clearance of cell-free DNA from plasma in relation to its methylation status, are poorly understood processes.. ...
Polyadenylation is among the most significant factors determining expression levels and is essential for the export of mRNA from the nucleus and subsequent translation, in addition to being a crucial component of mRNA stability (Ma et al., 2003). Tissue‐specific promoters, such as those expressed in cereal seeds, target the protein production to particular cells allowing easier harvesting of this product and preventing toxicity at the parent plant that may inhibit growth (Twyman et al., 2003). In actuality, with the discovery of a notary promoter, work was done to extract proteins in the nectar of a flower, which is chosen by bees and concentrated into honey (Breithaupt, 1999). Honey has the benefits of being composed of almost sugar and concentrating the protein, greatly easing the elimination procedure. If eukaryotic membrane proteins fail to be correctly expressed in yeasts, attention is usually drawn to higher eukaryotic hosts, e.g. insect cells, mammalian cells or cell-free expression ...
More rapid and economical methods of drug +____________________________________ screening are sought by the pharmaceutical industry due to the dramatic projected increase in the number of "druggable" targets. This project aims to develop such a method based on the use of novel tRNA-based fluorescent labeling of the N-terminus of proteins combined with affinity capillary electrophoresis (ACE). AmberGen will evaluate a series of monofunctional and bifunctional detection/affinity tags, which will be specifically incorporated at the N-terminus of proteins during their cell-free expression using specially prepared tRNA conjugates. Because of the ability to uniformly fluorescence label target proteins while preserving their activity, potential drug interactions can be screened using ACE combined with light induced fluorescence detection (LIF). In this approach, small alterations in mobility can be detected using low volumes of reagents. Photocleavable affinity tags provide a means to rapidly isolate ...
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... | → Synthelis is a French biotech company specialized in production, purification and characterization of membrane proteins. Discover our biotech blog.
Metabolic syndrome is directly linked with atherosclerotic burden and cell-free nucleic acids (cf-NA) analysis has recently emerged as a novel research tool in atherosclerosis prac..
We offer an assortment of used and refurbished Synthesizers/Sequencers ranging from DNA Sequencers, Protein Synthesizers, Peptide Synthesizers, and Bioanalyzers. Some of our popular manufacturers include Molecular Devices, ABI, Beckman and Agilent.
Introduction to chemical genomics / Paul R. Caron -- Chemistry for chemical genomics / Lutz Weber -- Computer-aided design of small molecules for chemical genomics / Philip M. Dean -- Design, synthesis, and screening of biomimetic ligands for affinity chromatography / Ana Cecília A. Roque, Geeta Gupta, and Christopher R. Lowe -- The role and application of in silico docking in chemical genomics research / Aldo Jongejan ... [et al.] -- Synthesis of complex carbohydrates and glyconjugates: enzymatic synthesis of globotetraose using #-1,3-N-acetylgalactosaminyltransferase LgtD from Haemophilus infuenzae strain Rd / Kang Ryu ... [et al.] -- High-throughput cloning for proteomics research / Sharon A. Doyle -- Screening for the expression of soluble recombinant protein in Escherichia coli / Sharon A. Doyle -- High-throughput purification of hexahistidine-tagged proteins expressed in Escherichia coli / Michael B. Murphy and Sharon A. Doyle -- The wheat germ cell-free expression system: methods for ...
... Starting to drench the goose eggs at the 14th day is for dissemination of embryo and eggshells brittle, complementary measures aimed at helping embryonic shells pecked. Without this system, it takes a lot of human resources and work hours to do the work. Use drenching free system in waterfowl incubator, adjust the micro-environment of incubator, did not affect hatching performance. Hatching rate of fertilized eggs could be enhanced 1% by the improvement and upgrading of equipment and technique. According to calculation, hatching operators hatches 100,000 goose eggs monthly, and the fertility rate is 80% and the hatching rate of fertilized eggs is also80%. If the gosling was 70 NTD for each, the production could increase income about 38,400 NTD for a month, and it could increase 460,800 NTD for a year; this result could be the reference for the industry.. (C. C. Hsiao, S. D. Wang and Y. S. Jea ...
Stage 14 Drosophila oocytes are arrested in first meiotic metaphase. A cell-free extract of these oocytes catalyzes apparent disassembly of purified Drosophila nuclei as well as of nuclear lamin polymers formed in vitro from isolated interphase lamins. Biochemically, the oocyte extract catalyzes lamin solubilization and phosphorylation as well as characteristic changes in one- and two-dimensional gel mobility. A previously unidentified soluble lamin isoform is easily seen after in vitro disassembly. This isoform is detectable but present only in very small quantities in vivo and is apparently derived specifically from one of the two interphase lamin isoforms. Cell-free nuclear lamina disassembly is ATP-dependent and addition of calcium to extracts blocks disassembly as judged both morphologically and biochemically. This system will allow enzymological characterization of cell-free lamina disassembly as well as molecular analysis of specific Drosophila mutants. ...
Background and Aims: Human embryonic stem cells (hESCs) are pluripotent cells and thus provide a promising cell source for clinical applications of regenerative medicine. Currently hESCs are cultured on fibroblast feeder cell layers, which provide necessary cell-cell interactions for the attachment and soluble factors enabling the undifferentiated growth of hESCs. However, culturing of feeder cells is expensive and laborious. In addition, xeno-products, used in feeder cell and hESC cultures could transmit animal pathogens to hESCs, and cause rejections when transplanted to patients. Therefore there is a need to develop xeno- and feeder cell-free culturing methods for hESCs. The first aim of this research project was to set up and compare two commercial xeno-products containing feeder cell-free culturing methods for hESCs. The second aim was to optimize a novel, defined, serum- and xeno-free Reges medium, developed in Regea, into feeder cell-free conditions ...
Using fluorescent and electron microscopy a comparative analysis was performed of components of the protein-synthesizing system of hippocampal neurons both in ground squirrels in various phases of the torpor-activity cycle and in rats cooled under the hypoxia-hypercapnia conditions. Results of the study have shown that in hippocampal neurons of the ground squirrels entering the natural torpor state and of rats under conditions of artificial hypothermia to 17°C, similar mechanisms might be possible to function, one of their obligatory components being a generalized decrease of activity of the protein-synthesizing system with its subsequent restoration at the exit from hypothermia. Cessation of hypoxia-hypercapnia even under conditions of a further temperature decrease restored the rat neuronal protein-synthesizing activity, which seems to indicate the presence of a potential possibility of adaptation of brain neurons in vivo to low temperatures, at which the integral organism of non-hibernating ...
AB - Cell-free transcription-translation systems were originally applied towards in vitro protein production. More recently, synthetic biology is enabling these systems to be used within a systematic design context for prototyping DNA regulatory elements, genetic logic circuits and biosynthetic pathways. The Gram-positive soil bacterium, Bacillus subtilis, is an established model organism of industrial importance. To this end, we developed several B. subtilis-based cell-free systems. Our improved B. subtilis WB800N-based system was capable of producing 0.8 µM GFP, which gave a ~72x fold-improvement when compared with a B. subtilis 168 cell-free system. Our improved system was applied towards the prototyping of a B. subtilis promoter library in which we engineered several promoters, derived from the wild-type Pgrac (σA) promoter, that display a range of comparable in vitro and in vivo transcriptional activities. Additionally, we demonstrate the cell-free characterisation of an inducible ...
Background- The ability to consistently detect cell-free tumor-specific DNA in peripheral blood of patients with metastatic breast cancer provides the opportunity to detect changes in tumor burden and to monitor response to treatment. Studies of cell-free DNA in the peripheral blood of breast cancer patients suggest that methylated DNA markers in serum or plasma could be used for detection of advanced disease, monitoring of therapeutic response, and for early detection of disease recurrence.. Methods- A genome-wide serum DNA methylome array (Illumina HumanMethylation27 BeadChip) analysis was conducted on cell-free circulating DNA in serum from women with stage IV recurrent breast cancer, and 232 key CpG loci were identified. Methylation for this panel of 10 gene loci was evaluated using our newly developed cMethDNA assay to detect miniscule amounts of methylated DNA in Training and Test sets of sera from a total of 112 women (n = 55 normal, n = 57 metastatic breast cancer). The clinical ...
A central goal of synthetic biology is to engineer cellular behavior by engineering synthetic gene networks for a variety of biotechnology and medical applications. The process of engineering gene networks often involves an iterative â design-build-testâ cycle, whereby the parts and connections that make up the network are built, characterized and varied until the desired network function is reached. Many advances have been made in the design and build portions of this cycle. However, the slow process of in vivo characterization of network function often limits the timescale of the testing step. Cell-free transcription-translation (TX-TL) systems offer a simple and fast alternative to performing these characterizations in cells. Here we provide an overview of a cell-free TX-TL system that utilizes the native Escherichia coli TX-TL machinery, thereby allowing a large repertoire of parts and networks to be characterized. As a way to demonstrate the utility of cell-free TX-TL, we illustrate the ...
The DNA replication (or origin) licensing machinery ensures precise duplication of the genome and contributes to the regulation of proliferative capacity in metazoa. Using an in vitro fibroblast model system coupled to a cell-free DNA replication assay, we have studied regulation of the origin licensing pathway during exit from and re-entry into the mitotic cell cycle. We show that in the quiescent state (G0) loss of proliferative capacity is achieved in part through down-regulation of the replication licensing factors Cdc6 and Mcm2-7. The origin licensing repressor geminin is absent in quiescent fibroblasts, suggesting that this powerful inhibitor of the licensing machinery is not required to suppress proliferative capacity in G0. Geminin expression is induced at a late stage in the G0-S transition post pre-RC assembly. Ectopic geminin can block re-acquisition of DNA replication competence during re-entry into the cell cycle, indicating that geminin levels must be tightly down-regulated for ...
Upregulation of antiapoptotic BCL2 family members is implicated in treatment-resistant lung cancers. The BCL2 homology 4 (BH4) domain is required for the antiapoptotic function of BCL2, leading Han and colleagues to seek a selective BCL2 BH4 antagonist for the treatment of lung cancer. The small-molecule BCL2 BH4 domain antagonist BDA-366 was identified as the most effective compound against human lung cancer cell lines, including both small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) cells. Sensitivity to BDA-366 was dependent on both BCL2 expression and BH4 binding, and BDA-366 selectively bound with high affinity to BCL2, but not to other BCL2 family members. In vitro cell-free assays demonstrated that BDA-366 induced a conformational change in BCL2 that exposed the BH3 domain, resulting in abrogation of its prosurvival function and conversion to a prodeath protein. BDA-366-induced apoptosis was mediated by BCL2-dependent BAX activation and cytochrome c release. In vivo ...
We set out to develop and apply a high-throughput cell-free protein synthesis (CFPS) platform that provides functional genomics information for a wide variety of open reading frames (ORFs). We then used this information to improve CFPS yields by 4- to 5-fold, depending on the protein product. With the increasing number of completed genome sequences and ongoing sequencing projects, the post-genomic era has ushered in the promise of complete understanding of biological systems. For such a task, the most important set of information is inarguably the knowledge of the function of each gene product. To lead this endeavor of discerning the properties and functions of the entirety of an organisms genes and gene products, the field of functional genomics has emerged. Functional genomics focuses on dynamic cellular aspects, such as gene transcription, translation, and protein-protein interactions, in attempts to understand the relationship between an organisms genome and its phenotype. Thus, the ...
Ricin A chain has been isolated from extremely toxic RCA II. Our ricin A chain has less than 0.01% of the toxicity of the native lectin in a cell culture test system, yet is as potent as native ricin in a cell-free protein synthesis assay.
ABCB10, or ATP binding cassette sub-family B member 10, is a protein localized in the mitochondrial inner membrane. It belongs to the ABC transporter family whose members are proteins that facilitate substrate transport across various biological membranes. It has been found that ABCB10 is required for normal heme biosynthesis during erythroid differentiation and also plays a role in protection against the damage caused by reactive oxygen species (ROS) production. This protective effect exists both in the erythrocyte development and in the heart recovery after the ischemia-reperfusion injury. However, as an ABC transporter, its transported substrates are not known, neither is the mechanism by which ABCB10 plays a role in protection against ROS damage. In this dissertation an 8-azido-ATP photolabeling system is established to study the ATP binding and hydrolysis properties of ABCB10. Using this approach, it is found that the conserved amino acid residues Gly497 and Lys498 in the Walker A motif of ...
The structure-activity relationships were determined for adrenergic compounds which either activated or blocked the activation of a partially purified adenyl cyclase isolated from frog erythrocytes. The results suggested that the presence of a β-hydroxyl group was essential for activity and that the potency of agonists as well as antagonists increased with the size of the substituent group of the amino nitrogen. In addition to the requirements for receptor affinity, compounds with intrinsic activity (agonists) had to have either OH or CH2OH substituents in both the m- and p-positions of the benzene ring. Since these structural requirements agreed well with those reported for intact tissue preparations, utilization of this relatively simple, cell-free preparation of adenyl cyclase may be a useful method for studying compounds with beta-adrenengic activity and for further defining the chemical nature of a beta-adrenergic receptor.. ...
A method for peptide mapping of tryptic digests on cellulose this layers was developed. The method was much more sensitive than procedures based on paper or column separation techniques, and enabled the detection of many peptides resulting from non-specific tryptic hydrolysis during protein digestion. An E-coli cell-free protein synthesizing system directed by R17 RNA was used to examine the fidelity of translation in vitro. The major product of this system, the coat protein, was separated from the in vitro reaction mixtures and analysed by the peptide mapping procedure. A series of experiments utilising radioactive amino acids established that the fidelity of translation was high, and enabled the identification of the mapping position of the major tryptic peptides ...
I started tfp when we got our above ground pool a few months ago. With a chronically ill child who also has severe eczema, I need help. Lil mans skin looks awful because he wants to swim daily when not at Drs. And if I skip a couple days of testing the pool water because were at these appointments, I come home to a green pool. Please help. Should I just bite the bullet and switch to one of the chlorine free systems everyone keeps pushing me towards? Im so confused.
For Quartering large bodies of armed troops among us: For protecting them, by a mock Trial, from punishment for any Murders which they should commit on the Inhabitants of these States: For cutting off our Trade with all parts of the world: For imposing Taxes on us without our Consent: For depriving us in many cases, of the benefits of Trial by Jury: For transporting us beyond Seas to be tried for pretended offences For abolishing the free System of English Laws in a neighbouring Province, establishing therein an Arbitrary government, and enlarging its Boundaries so as to render it at once an example and fit instrument for introducing the same absolute rule into these Colonies: For taking away our Charters, abolishing our most valuable Laws, and altering fundamentally the Forms of our Governments: For suspending our own Legislatures, and declaring themselves invested with power to legislate for us in all cases whatsoever. He has abdicated Government here, by declaring us out of his Protection and ...
Summary of Facts and Submissions. I. European patent No. 0 985 032 with the title: Ribosome complexes as selection particles for in vitro display and evolution of proteins was granted with 29 claims on the basis of the European application No. 98924468.6 corresponding to the international application No. PCT/GB98/01564 published as WO 98/054312.. II. An opposition was filed under Article 100(a) and (b) EPC for lack of novelty, inventive step and industrial application as well as for lack of a sufficient disclosure. The patent was maintained in amended form on the basis of a set of claims comprising claims 1 to 28 filed at oral proceedings.. Claim 1 read as follows:. 1. A method for the display and selection of proteins or peptides and for the recovery of the genetic material encoding them, which method consists of :. (a) transcription and translation of DNA in a eukaryotic cell free transcription/translation system such that complexed particles are formed, each comprising at least one ...
Circulating cell-free DNA (cfDNA) which may be extracted from plasma or serum by noninvasive procedures has demonstrated great potential to anticipate SL 0101-1 treatment response and survival for cancer patients. grouped regarding to genotype discovered in cfDNA. Nevertheless NSCLC sufferers which harbored activating mutation in cfDNA got a greater potential for response to EGFR-TKIs (chances proportion or OR 1.96 95 CI 1.59 No significant publication bias was discovered in this scholarly research. To conclude cfDNA could become a predictive and prognostic biomarker for sufferers with NSCLC. mutation position in NSCLC [17 18 These MMP17 evidences recommended that genotype in cfDNA is actually a guaranteeing tumor biomarker for NSCLC. A lot of studies got looked into the predictive or prognostic worth of cfDNA focus in NSCLC sufferers lately [19-22] (discover Table ?Desk11 for sources). These research were often little and SL 0101-1 reported various outcomes However. A few of them demonstrated a ...
Boone, C; Blackman, K; and Brandchaft, P, "Tumour immunity induced in mice with cell-free homogenates of influenza virus-infected tumour cells." (1971). Subject Strain Bibliography 1971. 390 ...
The final structure is NcoI-AccIII-...-SphI...-BamHI-XhoI. Now, when these coils are inserted into pBEST-Ptar-UTR1-NcoI_XhoI-T500. Now, the first polypeptide of a translational fusion can be inserted between NcoI//AccII and the second between BamHI//XhoI. Finally, SphI can be used to verify construction by recutting ...
Todays computer aided design systems enable the creation of digital product definitions that are widely used throughout the design process, for example in analysis or manufacturing. Typically, such product definitions are created after the bulk of [shape] designing has been completed because their creation requires a detailed knowledge of the shape that is to be defined. Consequently, there is a gulf between the exploration processes that result in the selection of a design concept and the creation of its definition. In order to address this distinction, between design exploration and product definition, understanding of how designers create and manipulate shapes is necessary. The research outlined in this paper results from work concerned with addressing these issues, with the long term goal of informing a new generation of computer aided design systems which support design exploration as well as the production of product definitions. This research is based on the shape grammar formalism ...
Subunit 9 of mitochondrial ATPase (Su9) is synthesized in reticulocyte lysates programmed with Neurospora poly A-RNA, and in a Neurospora cell free system as a precursor with a higher apparent molecular weight than the mature protein (Mr 16,400 vs. 10,500). The RNA which directs the synthesis of Su9 precursor is associated with free polysomes. The precursor occurs as a high molecular weight aggregate in the postribosomal supernatant of reticulocyte lysates. Transfer in vitro of the precursor into isolated mitochondria is demonstrated. This process includes the correct proteolytic cleavage of the precursor to the mature form. After transfer, the protein acquires the following properties of the assembled subunit: it is resistant to added protease, it is soluble in chloroform/methanol, and it can be immunoprecipitated with antibodies to F1-ATPase. The precursor to Su9 is also detected in intact cells after pulse labeling. Processing in vivo takes place posttranslationally. It is inhibited by the ...
University of Tennessee at Knoxville Doctoral student, Peter Golden Shankles, presents his dissertation on Interfacing to Biological Systems Using Microfluidics, discussing the popular new field of microfluidics and the 3D printed tools that are propelling it forward-for this project and numerous others recently too. 3D printing allows for much greater self-sustainability in the lab as researchers can create tools for experiments and chemical reactions on their own, but attention must also be paid to how such technologies affect chemical transformations and influence biological systems.. As microfluidics are used more frequently with cell-free protein synthesis systems (CFPS), researchers usually set up tubes to experiment with reactions. As the author points out, this is usually easy, but other studies have shown better success with engineered reaction hardware. In this study, the researchers aim to begin using microfluidics as well as nanoscale membranes to lessen distances in diffusion ...
TY - JOUR. T1 - Cell-free synthesis of enzymically active tissue-type plasminogen activator. T2 - Protein folding determines the extent of N-linked glycosylation. AU - Bulleid, N. J.. AU - Bassel-Duby, R. S.. AU - Freedman, R. B.. AU - Sambrook, J. F.. AU - Gething, M. J H. PY - 1992. Y1 - 1992. N2 - Tissue-type plasminogen activator (t-PA) is synthesized in mammalian cells as a mixture of two forms that differ in their extent of N-linked glycosylation. We have investigated the mechanism underlying this variation in glycosylation, using a cell-free system that consists of a rabbit reticulocyte lysate optimized for the formation of disulphide bonds and supplemented with dog pancreas microsomal membranes. Molecules of human t-PA synthesized in vitro are enzymically active and responsive to natural activators and inhibitors, and are glycosylated in a pattern identical with that of the protein produced in vivo. This demonstrates that t-PA synthesized in vitro folds into the same conformation as the ...
The identification of a Sir2-related enzyme in the mammalian mitochondrion raises a number of interesting questions related to the NAD-dependent enzymatic activity associated with this family of enzymes and the pivotal role played by NAD in mitochondrial metabolism. In almost every respect, hSIRT3 behaves as a classical mitochondrial matrix protein. Its dependency on an NH2-terminal cleavable presequence has been reported for other mitochondrial matrix proteins (Roise et al., 1986, 1988; von Heijne et al., 1989; Abe et al., 2000). Mitochondrial targeting sequences are characterized by the presence of positively charged and hydrophobic residues (negative charged residues are very rare) (Roise et al., 1988) and tend to adopt a helical, frequently amphipathic, conformation. Mutational analysis of an amphipathic helix within the NH2 terminus of hSIRT3 showed that eliminating the positive charges by substituting arginines with glycines or glutamines led to a loss of mitochondrial import. Disrupting ...
In the study to evaluate the anti-inflammatory potential of a series of trihydroxyflavones by testing their ability to scavenge reactive oxygen species (ROS) and reactive nitrogen species (RNS) in cells and cell-free systems and to inhibit the proinflammatory pathways mediated by the enzymes cyclooxygenase (COX) and 5-lipoxygenase (5-LOX), in which reactive species have a proven involvement, showed that The tested trihydroxyflavones proved to be effective inhibitors of neutrophils oxidative burst and were shown to scavenge different ROS and RNS in cell-free systems. The most active compound in the majority of the assays was 3,3,4-trihydroxyflavone, which was somehow expected due to the presence of the ortho-dihydroxy in the B-ring, an important structural feature in terms of free radical scavenging activity and the compounds were able to inhibit the production of leukotriene B(4) by 5-LOX in activated neutrophils. 3,5,7-Trihydroxyflavone was able to inhibit both COX-1 and COX-2, which makes ...
When programmed with yeast prepro-α-factor mRNA, the heterologous reticulocyte/dog pancreas translation system synthesizes two pheromone related polypeptides, a cytosolically located primary translation product (pp-α-Fcyt, 21 kDa) and a membrane-specific and multiply glycosylated e-factor precursor (pp-α-F3, 27.5 kDa). Glycosylation of the membrane specific pp-α-F3 species is competitively inhibited by synthetic peptides containing the consensus sequence Asn-Xaa-Thr as indicated by a shift of its molecular mass from 27.5 kDa to about 19.5 kDa (pp-α-F0), whereas the primary translation product pp-α-F cyt is not affected. Likewise, only the glycosylated pp-α-F3 structure is digested by Endo H yielding a polypeptide with a molecular mass between PP-α-F0 and pp-α-F cyt. These observations strongly suggest that the primary translation product is proteolytically processed during/on its translocation into the lumen of the microsomal vesicles. We believe that this proteolytic processing is due ...

Cell-free system - WikipediaCell-free system - Wikipedia

A cell-free system is an in vitro tool widely used to study biological reactions that happen within cells apart from a full ... where a cell-free translation system based on Escherichia coli (E. coli), of the cell extract-based type, had the mRNA template ... Cell-free systems may be divided into two primary classifications: cell extract-based, which remove components from within a ... Notably, in work leading to a Nobel prize the Nirenberg and Matthaei experiment used a cell-free system, of the cell extract- ...
more infohttps://en.wikipedia.org/wiki/Cell-free_system

James Swartz on Cell-Free Systems | AIChEJames Swartz on Cell-Free Systems | AIChE

... and get a preview of his upcoming talk at the Cell Free Systems Conference. ... Hear from James about promising career paths in cell-free systems, ... The Cell Free Systems Conference will focus on understanding, harnessing, and expanding the capabilities of biological systems ... What topic will you be addressing at the Cell Free Systems Conference?. I will describe how we are using the new process and ...
more infohttps://www.aiche.org/chenected/2019/10/james-swartz-on-cell-free-systems

CRISPRs Unwanted Edits Unmasked Using Faster Cell Free SystemCRISPR's Unwanted Edits Unmasked Using Faster Cell Free System

... "cell-free" system eliminated a lot of the complex biological activity within a cell that makes it hard to isolate the full ... CRISPRs Unwanted Edits Unmasked Using Faster Cell Free System. December 9, 2019. 0 ... it frees scientists to execute and screen multiple trial edits, and many more than is practical with a cell-based system. This ... which is pioneering advances in editing DNA plasmids extracted from human cells. The team has already has used the cell-free ...
more infohttps://www.genengnews.com/news/crisprs-unwanted-edits-unmasked-using-faster-cell-free-system/

Enhanced Protein Synthesis in a Cell-Free System from Hypertrophied Skeletal Muscle | ScienceEnhanced Protein Synthesis in a Cell-Free System from Hypertrophied Skeletal Muscle | Science

Enhanced Protein Synthesis in a Cell-Free System from Hypertrophied Skeletal Muscle ... Enhanced Protein Synthesis in a Cell-Free System from Hypertrophied Skeletal Muscle ... Enhanced Protein Synthesis in a Cell-Free System from Hypertrophied Skeletal Muscle ... Enhanced Protein Synthesis in a Cell-Free System from Hypertrophied Skeletal Muscle ...
more infohttp://science.sciencemag.org/content/157/3791/935

Human immunodeficiency virus integration in a cell-free system. | Journal of VirologyHuman immunodeficiency virus integration in a cell-free system. | Journal of Virology

Human immunodeficiency virus integration in a cell-free system.. V Ellison, H Abrams, T Roe, J Lifson, P Brown ... Human immunodeficiency virus integration in a cell-free system. Message Subject (Your Name) has forwarded a page to you from ... Integration of the viral genome into the nuclear DNA of a host cell plays a pivotal role in the replication of retroviruses. We ... integration by using extracts from HIV-infected cells. Analysis of the reaction products showed that HIV integration in vitro ...
more infohttps://jvi.asm.org/content/64/6/2711?ijkey=8a6824077b0857d4ad40a6b784b4b73f38a41bf0&keytype2=tf_ipsecsha

Cytokinesis signaling visualized in vitro, using Xenopus cell-free system
	Cytokinesis signaling visualized in vitro, using Xenopus cell-free system

Cytokinesis signaling visualized in vitro, using Xenopus cell-free system. Cytokinesis, when two daughter cells are physically ... have reconstituted the organization of cytokinesis signaling outside living cells, using a the cell-free Xenopus system. The ... We developed a cell-free system to recapitulate cytokinesis signaling using cytoplasmic extract from Xenopus eggs. Microtubules ... Spatial organization of cytokinesis signaling reconstituted in a cell-free system. Phuong A. Nguyen et al. Science 346, 244 ( ...
more infohttp://www.xenbase.org/entry/doNewsRead.do?id=167

Cell-free system for synthesizing membrane proteins cell free method for synthesizing membrane proteins (Patent) | DOepatentsCell-free system for synthesizing membrane proteins cell free method for synthesizing membrane proteins (Patent) | DOepatents

Title: Cell-free system for synthesizing membrane proteins cell free method for synthesizing membrane proteins. ... Cell-free system for synthesizing membrane proteins cell free method for synthesizing membrane proteins ... A low power EM sensor is used to detect the motions of windpipe tissues in the glottal region of the human speech system before ... A low power EM sensor is used to detect the motions of windpipe tissues in the glottal region of the human speech system before ...
more infohttps://www.osti.gov/doepatents/biblio/1084360-cell-free-system-synthesizing-membrane-proteins-cell-free-method-synthesizing-membrane-proteins

Susceptibility of influenza viruses to hypothiocyanite and hypoiodite produced by lactoperoxidase in a cell-free systemSusceptibility of influenza viruses to hypothiocyanite and hypoiodite produced by lactoperoxidase in a cell-free system

The results presented here show that the LPO/H2O2/(SCN-/I-) cell-free, in vitro experimental system is a functional tool to ... Our aim was to study the antiviral effect and substrate specificity of LPO to inactivate influenza viruses using a cell-free ... We propose that a LPO-based antiviral system is an important contributor to anti-influenza virus defense of the airways. ... These studies tested the hypothesis that influenza strains are all susceptible to the LPO-based antiviral system but exhibit ...
more infohttps://journals.plos.org/plosone/article?id=10.1371/journal.pone.0199167

Multi-Input Regulation and Logic with T7 Promoters in Cells and Cell-Free SystemsMulti-Input Regulation and Logic with T7 Promoters in Cells and Cell-Free Systems

... coli cell-free protein expression system. Our results expand the utility of T7 promoters in cell based as well as cell-free ... offer a simple orthogonal expression system for use in a variety of cellular backgrounds and even in cell-free systems. Here we ... two different transcriptional repressors for the construction of a logic gate that functions in cells and in cell-free systems ... Engineered gene circuits offer an opportunity to harness biological systems for biotechnological and biomedical applications. ...
more infohttp://journals.plos.org/plosone/article?id=10.1371/journal.pone.0078442

Biosynthesis of heparin : Modulation of polysaccharide chain length in a cell-free systemBiosynthesis of heparin : Modulation of polysaccharide chain length in a cell-free system

Biosynthesis of heparin: Modulation of polysaccharide chain length in a cell-free system. Lidholt, Kerstin Uppsala University, ...
more infohttp://uu.diva-portal.org/smash/record.jsf?pid=diva2:455588

Nod2-Nodosome in a Cell-Free System: Implications in Pathogenesis and Drug Discovery for Blau Syndrome and Early-Onset...Nod2-Nodosome in a Cell-Free System: Implications in Pathogenesis and Drug Discovery for Blau Syndrome and Early-Onset...

... which was suitable for wheat germ cell-free protein synthesis [20]. These results suggested that the wheat germ cell-free ... N. Kaneko, Y. Ito, T. Iwasaki et al., "Reconstituted AIM2 inflammasome in cell-free system," Journal of Immunological Methods, ... It is noted that there are some limitations of nodosome in a cell-free system. Only the initial event of Nod2-RICK interaction ... MDP-Induced Interaction between Nod2 and RICK in Nodosome in a Cell-Free System. Nod2-WT-Btn was coprecipitated with FLAG-RICK- ...
more infohttps://www.hindawi.com/journals/tswj/2016/2597376/

Linear DNA for rapid prototyping of synthetic biological circuits in an Escherichia coli based TX-TL cell-free systemLinear DNA for rapid prototyping of synthetic biological circuits in an Escherichia coli based TX-TL cell-free system

... Zachary ... protect linear DNA strands from exonuclease degradation in an Escherichia coli based transcription-translation cell-free system ...
more infohttp://www.cds.caltech.edu/~murray/papers/sun+13-acs_synbio.html

Molecules | Free Full-Text | Antioxidant Activity of Ixora parviflora in a Cell/Cell-Free System and in UV-Exposed Human...Molecules | Free Full-Text | Antioxidant Activity of Ixora parviflora in a Cell/Cell-Free System and in UV-Exposed Human...

The aim of this study was to investigate the antioxidant activity of Ixora parviflora extract (IPE) in a cell-free system and ... Antioxidant Activity of Ixora parviflora in a Cell/Cell-Free System and in UV-Exposed Human Fibroblasts. Kuo-Ching Wen 1, Hua- ... "Antioxidant Activity of Ixora parviflora in a Cell/Cell-Free System and in UV-Exposed Human Fibroblasts." Molecules 16, no. 7: ... Antioxidant Activity of Ixora parviflora in a Cell/Cell-Free System and in UV-Exposed Human Fibroblasts. Molecules 2011, 16, ...
more infohttp://www.mdpi.com/1420-3049/16/7/5735

Difference between revisions of Establishing microfluidic cell-free systems for the rapid prototyping of synthetic genetic...Difference between revisions of "Establishing microfluidic cell-free systems for the rapid prototyping of synthetic genetic...

This study is designed to demonstrate that biological systems can be forward and reverse engineered in cell-free environments. ... Difference between revisions of "Establishing microfluidic cell-free systems for the rapid prototyping of synthetic genetic ... cell-free_systems_for_the_rapid_prototyping_of_synthetic_genetic_networks&oldid=22493" ... We propose to develop a closed feedback system that controls the microfluidic system, runs experiments and analyses results to ...
more infohttp://www.cds.caltech.edu/~murray/wiki/index.php?title=Establishing_microfluidic_cell-free_systems_for_the_rapid_prototyping_of_synthetic_genetic_networks&diff=22493&oldid=18429

Frontiers | Wheat germ cell-free system-based production of hemagglutinin-neuraminidase glycoprotein of human parainfluenza...Frontiers | Wheat germ cell-free system-based production of hemagglutinin-neuraminidase glycoprotein of human parainfluenza...

A recombinant full-length HPIV3-HN was successfully synthesized using the wheat-germ cell-free protein production system. After ... After immunization and cell fusion, 36 mouse hybridomas producing MAbs to HPIV3-HN were established. The MAbs obtained were ... immunization and cell fusion, 36 mouse hybridomas producing MAbs to HPIV3-HN were established. The MAbs obtained were fully ... A recombinant full-length HPIV3-HN was successfully synthesized using the wheat-germ cell-free protein production system. ...
more infohttps://www.frontiersin.org/articles/10.3389/fmicb.2014.00208/full

Disassembly of the Drosophila nuclear lamina in a homologous cell-free system | Journal of Cell ScienceDisassembly of the Drosophila nuclear lamina in a homologous cell-free system | Journal of Cell Science

This system will allow enzymological characterization of cell-free lamina disassembly as well as molecular analysis of specific ... Disassembly of the Drosophila nuclear lamina in a homologous cell-free system ... Disassembly of the Drosophila nuclear lamina in a homologous cell-free system ... Disassembly of the Drosophila nuclear lamina in a homologous cell-free system ...
more infohttp://jcs.biologists.org/content/108/5/2027

Lirias: The wheat germ cell-free system possesses processing activity for the precursor of human placental lactogenLirias: The wheat germ cell-free system possesses processing activity for the precursor of human placental lactogen

5. The presence of one or several processing enzymes in the wheat germ cell-free system was indicated by the effect of Triton X ... The wheat germ cell-free system possesses processing activity for the precursor of human placental lactogen. ... The poly(A)-containing fraction stimulated amino acid incorporation 5- to 10-fold in the wheat germ cell-free system. ... Education Domains Department of Molecular Cell Biology (-) › Biochemistry Section (Medicine) (-) › ...
more infohttps://lirias.kuleuven.be/handle/123456789/11410

KAKEN - Research Projects | Study on initiation of DNA repication in cell-free system of Xenopus eggs (KAKENHI-PROJECT-07680762)KAKEN - Research Projects | Study on initiation of DNA repication in cell-free system of Xenopus eggs (KAKENHI-PROJECT-07680762)

Study on initiation of DNA repication in cell-free system of Xenopus eggs. Research Project ... which has an activity to untwist DNA and open double strand DNA to single strand DNA from cell-free DNA synthetic system of ... Publications] Someya, A.: The association and involvement of the P1 protein family in a cell free extract of Xenopus eggs ... Publications] Someya.A.: The possible involvement of replication-related proteins with a DEAD-box-like motif in cell-free DNA ...
more infohttps://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07680762/

An ABCB10 cell-free system and the exploration of its substrates and regulatorsAn ABCB10 cell-free system and the exploration of its substrates and regulators

... ... In this dissertation an 8-azido-ATP photolabeling system is established to study the ATP binding and hydrolysis properties of ... Therefore, the 8-azido-ATP photolabeling system can be utilized to test potential substrates of ABCB10. Substances related to ... The 8-azido-ATP photolabeling system shows that GSSG stimulates ATP hydrolysis without affecting ATP binding, whereas GSH ...
more infohttps://open.bu.edu/handle/2144/15368

Functional expression of a single-chain antibody to ErbB-2 in plants and cell-free systems - Galeffi, Patrizia, Lombardi,...Functional expression of a single-chain antibody to ErbB-2 in plants and cell-free systems - Galeffi, Patrizia, Lombardi,...

... but a worldwide deficit in the manufacturing capacities of mammalian cell ... Functional expression of a single-chain antibody to ErbB-2 in plants and cell-free systems : Aberrant signaling by ErbB-2 (HER ... Research Open Access Functional expression of a single-cha in antibody to ErbB-2 in plants and cell-free systems Patrizia ... Functional expression of a single-chain antibody to ErbB-2 in plants and cell-free systems Galeffi, Patrizia, Lombardi, Alessio ...
more infohttps://www.youscribe.com/catalogue/documents/savoirs/functional-expression-of-a-single-chain-antibody-to-erbb-2-in-plants-2059553

Translation of homologous and heterologous messenger RNAs in a yeast cell-free system<...Translation of homologous and heterologous messenger RNAs in a yeast cell-free system<...

Hussain, I., & Leibowitz, M. J. (1986). Translation of homologous and heterologous messenger RNAs in a yeast cell-free system. ... Hussain, I & Leibowitz, MJ 1986, Translation of homologous and heterologous messenger RNAs in a yeast cell-free system, Gene ... Translation of homologous and heterologous messenger RNAs in a yeast cell-free system. In: Gene. 1986 ; Vol. 46, No. 1. pp. 13- ... Translation of homologous and heterologous messenger RNAs in a yeast cell-free system. / Hussain, Iffat; Leibowitz, Michael J. ...
more infohttps://ucdavis.pure.elsevier.com/en/publications/translation-of-homologous-and-heterologous-messenger-rnas-in-a-ye

Cytoplasmic components of apoptosis by irofulven revealed by cell-free apoptotic systems | Cancer ResearchCytoplasmic components of apoptosis by irofulven revealed by cell-free apoptotic systems | Cancer Research

Cytoplasmic components of apoptosis by irofulven revealed by cell-free apoptotic systems. Jan M. Woynarowski, Barbara A. ... Cytoplasmic components of apoptosis by irofulven revealed by cell-free apoptotic systems ... To better understand the nature of the differential responses of tumor and normal cells, we used a cell-free apoptosis ... Cytoplasmic components of apoptosis by irofulven revealed by cell-free apoptotic systems ...
more infohttp://cancerres.aacrjournals.org/content/64/7_Supplement/1195.4

Recombination between adenovirus type 12 DNA and a hamster preinsertion sequence in a cell-free system. Patch homologies and...Recombination between adenovirus type 12 DNA and a hamster preinsertion sequence in a cell-free system. Patch homologies and...

We have previously described a cell-free recombination system derived from hamster cell nuclear extracts in which the in vitro ... Further work will be required to ascertain that the cell-free recombination system mimics certain elements of the mechanisms of ... Recombination between adenovirus type 12 DNA and a hamster preinsertion sequence in a cell-free system. Patch homologies and ... Recombination between adenovirus type 12 DNA and a hamster preinsertion sequence in a cell-free system. Patch homologies and ...
more infohttp://www.biochemie.abi.med.uni-muenchen.de/publications/article/pub_1992_007.html
  • Attempts to create selective, laser-made chromosome breaks in mitotic mammalian cells have shown that unless the breakage occurs directly in the kinetochore region of the mitotic chromosome, there is no ATM and/or ATR dependent DNA damage checkpoint response that would inhibit mitosis progression [ 12 ]. (biomedcentral.com)
  • These systems have enabled cell-free synthetic biology to emerge, providing control over what reaction is being examined, as well as its yield, and lessening the considerations otherwise invoked when working with more sensitive live cells. (wikipedia.org)
  • In 1998, he moved to Stanford University as a Professor of Chemical Engineering focusing on cell-free biology. (aiche.org)
  • We've just published our special issue focussing on plant cell biology - check out the table of contents here , and a selection of the issue's highlights below. (biologists.org)
  • In this interview , our Special Issue's guest editor Jenny Russinova discusses her excitement for plant cell biology and what she hopes to achieve with the issue. (biologists.org)
  • We are now accepting submissions for our upcoming special issue on 'Reconstituting cell biology', guest edited by Manuel Théry. (biologists.org)
  • Cell-free systems are widely used for fundamental molecular biology, for applications in genetic engineering, in metabolic engineering or in medical diagnostics and for a better understanding in the origin of life 1,2,3 . (nature.com)
  • Cell-free systems present many advantages for synthetic biology through high-throughput characterization and prototyping of natural and synthetic circuits. (nature.com)
  • As our team is composed of a dry lab skilled in machine learning and a wet lab with experience in cell-free approach, it was natural to tackle this problem using the recent advance made in machine learning and in automation of biology. (nature.com)
  • The reduced complexity of protein production in cell-free expression systems results in a frequent correlation of efficiency problems with the essential transcription/translation process. (jascoinc.com)
  • The generated pool of DNA templates is analyzed in a cell-free expression screen and the most efficient template is selected for further preparative scale protein production. (jascoinc.com)
  • Even with the same procedure, the homemade preparation of the cell-free mix led to variability of the protein production efficiency. (nature.com)
  • We examined homologous and heterologous mixes of nuclei and cytosols from cancer cells (CEM and LNCaP-Pro5), which initiate apoptosis in response to irofulven, and from normal cells (NCM 460), which display a non-apoptotic cytostatic response. (aacrjournals.org)
  • Integration of the viral genome into the nuclear DNA of a host cell plays a pivotal role in the replication of retroviruses. (asm.org)
  • However, the preincubation of cytosols from normal cells with the drug had marginal effects on the nuclear morphology. (aacrjournals.org)
  • The potent induction of apoptosis in various types of tumor cells, contrasted to largely reversible non-apoptotic effects in normal cells is one of the hallmarks of a novel dual-action DNA- and protein-alkylating drug, irofulven (hydroxymethylacylfulvene). (aacrjournals.org)
  • The patchy homologies are similar to those found earlier during the analyses of some of the junction sequences in integrated Ad12 genomes in Ad12-induced hamster tumor cell lines. (uni-muenchen.de)
  • These findings demonstrate that irofulven-induced pro-apoptotic changes in cancer cells involve cytosolic factors. (aacrjournals.org)
  • Publications] Yokoyama, Y.: 'Enhancement of apoptosis in developing chick neural retina cells by basic fibroblast growth factor' Jounal of Neurochemistry. (nii.ac.jp)
  • Publications] Yokoyama, Y.: 'CPP32 activation during dolichyl phosphate-induced apoptosis in U937 leukemia cells' FEBS Letters. (nii.ac.jp)
  • These changes were observed with nuclei from both apoptosis-prone cancer cells and apoptosis-resistant normal cells. (aacrjournals.org)
  • In addition, exposure of cytosols from untreated cancer cells to irofulven (prior to incubation with nuclei) resulted in apoptosis in both cancer and normal nuclei. (aacrjournals.org)
  • The possibility that cytoplasmic factors influence cell sensitivity to apoptosis is consistent with our hypothesis that apoptotic effects of irofulven originate not only from DNA damage but also from protein damage. (aacrjournals.org)
  • As a central component of the cadherin adhesion complex, β-catenin is expressed widely in the cells of vertebrates. (rupress.org)
  • The aim of this study was to investigate the antioxidant activity of Ixora parviflora extract (IPE) in a cell-free system and erythrocytes, and the ability of IPE to inhibit reactive oxygen species (ROS) generation in human fibroblasts (Hs68) after ultraviolet (UV) exposure. (mdpi.com)
  • Chiang, H.-M. Antioxidant Activity of Ixora parviflora in a Cell/Cell-Free System and in UV-Exposed Human Fibroblasts. (mdpi.com)
  • Wen K-C, Chiu H-H, Fan P-C, Chen C-W, Wu S-M, Chang J-H, Chiang H-M. Antioxidant Activity of Ixora parviflora in a Cell/Cell-Free System and in UV-Exposed Human Fibroblasts. (mdpi.com)
  • A Travelling Fellowship from Journal of Cell Science allowed her to spend time in Prof Maddy Parson's lab at King's College London, learning new cell migration assays and analysing fibroblasts cultured from individuals with Parkinson's. (biologists.org)
  • We combined the predictive power of machine learning and the large-scale exploration of cell-free compositions offer by automation to maximize and predict protein productions in homemade cell-free systems. (nature.com)
  • A stable mRNA-dependent cell-free translation system from Saccharomyces cerevisiae, prepared by a modification of the method of Hofbauer et al. (elsevier.com)
  • Recently this system has been successfully used to uncover an ATM and ATR dependent checkpoint affecting centrosome driven spindle assembly. (biomedcentral.com)
  • There was a time dependent increase in the protein yield as well as the intensity of the native tetramer band in insect cell derived microsomes. (nanion.de)
  • Immunobloting exper iments using antibody against the 87 kDa polypeptide showed that behavior of DUF during a cell cycle or embryogenesis co-relates with DNA replication. (nii.ac.jp)
  • As results, we detected a protein complex composed of 5 prteeins containing MCM famliy which regulates intiaton of DNA replication, correspondiing too the license factor which allows DNA replication only once in a cell cycle. (nii.ac.jp)
  • To reduce the cost and power consumption of such systems, it is important to utilize low-quality transceiver hardware at the APs. (diva-portal.org)
  • Methods Herein, we describe a multi-platform approach for the production of recombinant Single chain Fragments of antibody variable regions (ScFvs) to ErbB-2 that involves their functional expression in (a) bacteria, (b) transient as well as stable transgenic tobacco plants, and (c) a newly developed cell-free transcription-translation system. (youscribe.com)
  • Nanion is pleased to announce the launch of Dynamite 8 , a Dynamic Clamp unit for the multichannel automated patch clamp system, the Patchliner. (nanion.de)
  • The Dynamite 8 simulates ion channel currents in cells in real-time during patch clamp experiments. (nanion.de)
  • Many of the studies analyzing the effects on mitosis of spindle poisoning or DNA damage inducing drugs have been performed using microscopy-based techniques monitoring the behavior of few cells or applying population-based approaches. (biomedcentral.com)
  • The complete process takes only few days and the synthesized PCR fragments can be used directly as templates for preparative scale cell-free reactions. (jascoinc.com)
  • In striking contrast, cytosols from irofulven-treated normal NCM 460 cells did not change the morphology of nuclei from any of the three cell lines. (aacrjournals.org)
  • A major stumbling block to probing the underlying mechanisms has been the lack of a cell-free and fully controllable experimental system. (xenbase.org)
  • Further work will be required to ascertain that the cell-free recombination system mimics certain elements of the mechanisms of integrative recombination and to purify the cellular components essential for recombination. (uni-muenchen.de)
  • Prion aggregates are extremely stable and accumulate in infected tissue, causing tissue damage and cell death. (wikipedia.org)
  • It is also very exciting to know that at least two cell-free produced biopharmaceuticals are in clinical trials and to also see the great progress in technologies for on-demand biopharmaceutical manufacturing. (aiche.org)
  • Just last month CRISPR Therapeutics and Vertex Pharmaceuticals announced positive safety and efficacy data from the first two patients enrolled in two clinical trials evaluating the CRISPR-Cas9 gene editing therapy CTX001, for sickle cell disease and beta thalassemia. (genengnews.com)
  • Nod2 was reported to be oligomerized with adaptor protein RICK (RIP2/RIPK2) and IKK complexes, which can activate NF- κ B by muramyl dipeptide (N-Acetylmuramyl-L-Alanyl-D-Isoglutamine: MDP), one of the components of bacterial cell-wall peptidoglycan, and is utilized as an immune-stimulatory adjuvant for vaccination and for developing antibodies [ 4 - 8 ]. (hindawi.com)
  • HN can be recognized by the host immune system and antibodies against epitopes within HN can neutralize its activity through the inhibition of the function and/or activity of either hemagglutinin or neuraminidase. (frontiersin.org)
  • Publications] Takayanagi, S.: 'Sulfolobus hakonensis sp.nov.,anovel species of acidothermophilic Archaeon' Int.J.System.Bacteriol.46・2. (nii.ac.jp)