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)
The biosynthesis of PEPTIDES and PROTEINS on RIBOSOMES, directed by MESSENGER RNA, via TRANSFER RNA that is charged with standard proteinogenic AMINO ACIDS.
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).
The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.
Established cell cultures that have the potential to propagate indefinitely.
The rate dynamics in chemical or physical systems.
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 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.
The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.
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 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.

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)

A cell-free system is a biochemical environment in which biological reactions can occur outside of an intact living cell. These systems are often used to study specific cellular processes or pathways, as they allow researchers to control and manipulate the conditions in which the reactions take place. In a cell-free system, the necessary enzymes, substrates, and cofactors for a particular reaction are provided in a test tube or other container, rather than within a whole cell.

Cell-free systems can be derived from various sources, including bacteria, yeast, and mammalian cells. They can be used to study a wide range of cellular processes, such as transcription, translation, protein folding, and metabolism. For example, a cell-free system might be used to express and purify a specific protein, or to investigate the regulation of a particular metabolic pathway.

One advantage of using cell-free systems is that they can provide valuable insights into the mechanisms of cellular processes without the need for time-consuming and resource-intensive cell culture or genetic manipulation. Additionally, because cell-free systems are not constrained by the limitations of a whole cell, they offer greater flexibility in terms of reaction conditions and the ability to study complex or transient interactions between biological molecules.

Overall, cell-free systems are an important tool in molecular biology and biochemistry, providing researchers with a versatile and powerful means of investigating the fundamental processes that underlie life at the cellular level.

Protein biosynthesis is the process by which cells generate new proteins. It involves two major steps: transcription and translation. Transcription is the process of creating a complementary RNA copy of a sequence of DNA. This RNA copy, or messenger RNA (mRNA), carries the genetic information to the site of protein synthesis, the ribosome. During translation, the mRNA is read by transfer RNA (tRNA) molecules, which bring specific amino acids to the ribosome based on the sequence of nucleotides in the mRNA. The ribosome then links these amino acids together in the correct order to form a polypeptide chain, which may then fold into a functional protein. Protein biosynthesis is essential for the growth and maintenance of all living organisms.

Deoxyribonucleic acid (DNA) is the genetic material present in the cells of organisms where it is responsible for the storage and transmission of hereditary information. DNA is a long molecule that consists of two strands coiled together to form a double helix. Each strand is made up of a series of four nucleotide bases - adenine (A), guanine (G), cytosine (C), and thymine (T) - that are linked together by phosphate and sugar groups. The sequence of these bases along the length of the molecule encodes genetic information, with A always pairing with T and C always pairing with G. This base-pairing allows for the replication and transcription of DNA, which are essential processes in the functioning and reproduction of all living organisms.

A base sequence in the context of molecular biology refers to the specific order of nucleotides in a DNA or RNA molecule. In DNA, these nucleotides are adenine (A), guanine (G), cytosine (C), and thymine (T). In RNA, uracil (U) takes the place of thymine. The base sequence contains genetic information that is transcribed into RNA and ultimately translated into proteins. It is the exact order of these bases that determines the genetic code and thus the function of the DNA or RNA molecule.

A cell line is a culture of cells that are grown in a laboratory for use in research. These cells are usually taken from a single cell or group of cells, and they are able to divide and grow continuously in the lab. Cell lines can come from many different sources, including animals, plants, and humans. They are often used in scientific research to study cellular processes, disease mechanisms, and to test new drugs or treatments. Some common types of human cell lines include HeLa cells (which come from a cancer patient named Henrietta Lacks), HEK293 cells (which come from embryonic kidney cells), and HUVEC cells (which come from umbilical vein endothelial cells). It is important to note that cell lines are not the same as primary cells, which are cells that are taken directly from a living organism and have not been grown in the lab.

In the context of medicine and pharmacology, "kinetics" refers to the study of how a drug moves throughout the body, including its absorption, distribution, metabolism, and excretion (often abbreviated as ADME). This field is called "pharmacokinetics."

1. Absorption: This is the process of a drug moving from its site of administration into the bloodstream. Factors such as the route of administration (e.g., oral, intravenous, etc.), formulation, and individual physiological differences can affect absorption.

2. Distribution: Once a drug is in the bloodstream, it gets distributed throughout the body to various tissues and organs. This process is influenced by factors like blood flow, protein binding, and lipid solubility of the drug.

3. Metabolism: Drugs are often chemically modified in the body, typically in the liver, through processes known as metabolism. These changes can lead to the formation of active or inactive metabolites, which may then be further distributed, excreted, or undergo additional metabolic transformations.

4. Excretion: This is the process by which drugs and their metabolites are eliminated from the body, primarily through the kidneys (urine) and the liver (bile).

Understanding the kinetics of a drug is crucial for determining its optimal dosing regimen, potential interactions with other medications or foods, and any necessary adjustments for special populations like pediatric or geriatric patients, or those with impaired renal or hepatic function.

Molecular sequence data refers to the specific arrangement of molecules, most commonly nucleotides in DNA or RNA, or amino acids in proteins, that make up a biological macromolecule. This data is generated through laboratory techniques such as sequencing, and provides information about the exact order of the constituent molecules. This data is crucial in various fields of biology, including genetics, evolution, and molecular biology, allowing for comparisons between different organisms, identification of genetic variations, and studies of gene function and regulation.

Messenger RNA (mRNA) is a type of RNA (ribonucleic acid) that carries genetic information copied from DNA in the form of a series of three-base code "words," each of which specifies a particular amino acid. This information is used by the cell's machinery to construct proteins, a process known as translation. After being transcribed from DNA, mRNA travels out of the nucleus to the ribosomes in the cytoplasm where protein synthesis occurs. Once the protein has been synthesized, the mRNA may be degraded and recycled. Post-transcriptional modifications can also occur to mRNA, such as alternative splicing and addition of a 5' cap and a poly(A) tail, which can affect its stability, localization, and translation efficiency.

Genetic transcription is the process by which the information in a strand of DNA is used to create a complementary RNA molecule. This process is the first step in gene expression, where the genetic code in DNA is converted into a form that can be used to produce proteins or functional RNAs.

During transcription, an enzyme called RNA polymerase binds to the DNA template strand and reads the sequence of nucleotide bases. As it moves along the template, it adds complementary RNA nucleotides to the growing RNA chain, creating a single-stranded RNA molecule that is complementary to the DNA template strand. Once transcription is complete, the RNA molecule may undergo further processing before it can be translated into protein or perform its functional role in the cell.

Transcription can be either "constitutive" or "regulated." Constitutive transcription occurs at a relatively constant rate and produces essential proteins that are required for basic cellular functions. Regulated transcription, on the other hand, is subject to control by various intracellular and extracellular signals, allowing cells to respond to changing environmental conditions or developmental cues.

'Escherichia coli' (E. coli) is a type of gram-negative, facultatively anaerobic, rod-shaped bacterium that commonly inhabits the intestinal tract of humans and warm-blooded animals. It is a member of the family Enterobacteriaceae and one of the most well-studied prokaryotic model organisms in molecular biology.

While most E. coli strains are harmless and even beneficial to their hosts, some serotypes can cause various forms of gastrointestinal and extraintestinal illnesses in humans and animals. These pathogenic strains possess virulence factors that enable them to colonize and damage host tissues, leading to diseases such as diarrhea, urinary tract infections, pneumonia, and sepsis.

E. coli is a versatile organism with remarkable genetic diversity, which allows it to adapt to various environmental niches. It can be found in water, soil, food, and various man-made environments, making it an essential indicator of fecal contamination and a common cause of foodborne illnesses. The study of E. coli has contributed significantly to our understanding of fundamental biological processes, including DNA replication, gene regulation, and protein synthesis.

"Cells, cultured" is a medical term that refers to cells that have been removed from an organism and grown in controlled laboratory conditions outside of the body. This process is called cell culture and it allows scientists to study cells in a more controlled and accessible environment than they would have inside the body. Cultured cells can be derived from a variety of sources, including tissues, organs, or fluids from humans, animals, or cell lines that have been previously established in the laboratory.

Cell culture involves several steps, including isolation of the cells from the tissue, purification and characterization of the cells, and maintenance of the cells in appropriate growth conditions. The cells are typically grown in specialized media that contain nutrients, growth factors, and other components necessary for their survival and proliferation. Cultured cells can be used for a variety of purposes, including basic research, drug development and testing, and production of biological products such as vaccines and gene therapies.

It is important to note that cultured cells may behave differently than they do in the body, and results obtained from cell culture studies may not always translate directly to human physiology or disease. Therefore, it is essential to validate findings from cell culture experiments using additional models and ultimately in clinical trials involving human subjects.

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- ...
... 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 ...
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Establishment of cell-free protein synthesis system for practical usage. 2004 - Atushi Miyawaki. The multidisciplinary ... Creation of cell sheet engineering based on intelligent surfaces. 2010 - Teruhiko Wakayama [ja]. Development of novel animal ... The research and development of Dy free Nd-Fe-B anisotropic bonded magnet and its applications to motors. 2013 - Hiroshi Harada ... High-level synthesis and verification on system LSI. 2005 - Mitsuo Usami, Ryo Imura. Development of ultra-small chip for IC ...
... is a Japanese biochemist and professor well known for developing the cell-free protein expression system based on ... Endo, Yaeta (11 May 2021). "Development of a cell-free protein synthesis system for practical use". Proceedings of the Japan ... Sawasaki, Tatsuya; Ogasawara, Tomio; Morishita, Ryo; Endo, Yaeta (12 November 2002). "A cell-free protein synthesis system for ... Madono, Masaki; Sawasaki, Tatsuya; Morishita, Ryo; Endo, Yaeta (April 2011). "Wheat germ cell-free protein production system ...
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1979-1980: The development of cell-free systems leads to the identification of complex sets of proteins called accessory ... 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 ...
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It can be expressed efficiently by a cell-free protein expression system. This enzyme belongs to the family of transferases, ... "Preparative scale cell-free production and quality optimization of MraY homologues in different expression modes". The Journal ... Higashi Y, Strominger JL, Sweeley CC (June 1967). "Structure of a lipid intermediate in cell wall peptidoglycan synthesis: a ...
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Appanna DL, Grundy BJ, Szczepan EW, Viswanatha T (1984). "Aerobactin synthesis in a cell-free system of Aerobacter aerogenes 62 ...
LAMFROM H. Factors determining the specificity of hemoglobin synthesized in a cell-free system. J Mol Biol. 1961 Jun;3:241-52. ... Lamfrom, Hildegard (1961-06-01). "Factors determining the specificity of hemoglobin synthesized in a cell-free system". Journal ... "Factors determining the specificity of hemoglobin synthesized in a cell-free system". Journal of Molecular Biology. 3 (3): 241- ... in a cell-free context. This allowed her to make a number of contributions to the field including providing some of the first ...
Gruenberg, J.; Howell, K. E. (1988). "Fusion in the endocytic pathway reconstituted in a cell-free system using immuno-isolated ... Using innovative approaches such as phospholipid-specific antibodies and reconstituted cell-free systems, Jean Gruenberg and ... "Reconstitution of vesicle fusions occurring in endocytosis with a cell-free system". The EMBO Journal. 5 (12): 3091-3101. doi: ... and motor-dependent fusion in vitro between apical and basolateral endocytic vesicles from MDCK cells". Cell. 62 (4): 719-731. ...
"Functional protein expression from a DNA based wheat germ cell-free system". J. Struct. Funct. Genomics. 8 (4): 199-208. doi: ... Promega was an early supplier in the cell-free protein synthesis field and is continuing to develop its portfolio in this area ... The company also sells their own Maxwell RSC and Maxwell RSC 48 Systems, bench-top automated purification systems for low and ... The Promega PowerPlex STR systems were the first commercially available systems for STR analysis that contained all of the ...
Chen CM, Melitz DK (1979). "Cytokinin biosynthesis in a cell-free system from cytokinin-autotrophic tobacco tissue cultures". ...
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 ...
Reduction of aromatic acids to aldehydes and alcohols in the cell-free system. 2. Purification and properties of aryl-alcohol: ...
Jinnai H, Nakamura S (August 1999). "Characterization of phospholipase D activation by GM2 activator in a cell-free system". ... The GPI-anchor is a glycolipid found on many blood cells. The protein encoded by the GPLD1 gene is a GPI degrading enzyme that ... Xiaotong H, Hannocks MJ, Hampson I, Brunner G (2002). "GPI-specific phospholipase D mRNA expression in tumor cells of different ... April 1994). "Release of GPI-anchored membrane proteins by a cell-associated GPI-specific phospholipase D". The EMBO Journal. ...
Gross GG, Zenk MH (1969). "[Reduction of aromatic acids to aldehydes and alcohols in the cell-free system. 1. Purification and ...
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, Lukas ... Cell. 23 (3): 307-18. doi:10.1016/j.molcel.2006.06.016. PMID 16885021. Peschiaroli A, Dorrello NV, Guardavaccaro D, et al. ( ...
Eukaryotic cell extracts may also be used in other cell-free systems, for example, the wheat germ cell-free expression systems ... Brödel AK, Wüstenhagen DA, Kubick S (2015). "Cell-Free Protein Synthesis Systems Derived from Cultured Mammalian Cells". ... Mammalian cell-free systems have also been produced. Expression vector in an expression host is now the usual method used in ... Wheat Germ Cell-Free Expression System for Protein Production". Current Protocols in Protein Science. Vol. Chapter 5. pp. 5.18. ...
Rohmer M, Anding C, Ourisson G (December 1980). "Non-specific biosynthesis of hopane triterpenes by a cell-free system from ... These cold- and desiccation-resistant cell structures are dormant and therefore not photosynthetically active, further ... Welander PV (August 2019). "Deciphering the evolutionary history of microbial cyclic triterpenoids". Free Radical Biology & ... methods and systems", issued 2016-10-19, assigned to California Institute of Technology (Articles with short description, Short ...
"A Ubiquitous Amino Acid Source for Prokaryotic and Eukaryotic Cell-Free Transcription-Translation Systems". Frontiers in ... leaving many smaller peptide chains alongside the free amino acids. Casamino acid is used as a component of microbiological ...
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- ...
... including the central nervous system (CNS). They are secreted by a range of cell types and via blood reaching other cells whose ... focusing on cell-cell communication in physiology and pathology. ... They are involved in cell differentiation, tissue homeostasis, ... we discuss what is known about the role and potential future applications of exosomes in the nervous system and its diseases, ... are membranous particles released by cells into the extracellular space. ...
NEBExpress® Cell-free E. coli Protein Synthesis System Applications:. NEBExpress® Cell-free E. coli Protein Synthesis System. ... Cell-free E. coli Protein Synthesis System (NEB #E5360). Using a positive control template to verify protein synthesis can be ... Cell-free E. coli Protein Synthesis System (NEB #E5360) ... Free Shipping. Save time and money by placing an order with NEB ... Take advantage of free shipping for any order totaling over $350. Place your order before 7:30pm EST for overnight delivery. ...
Probing metabolism in an E. coli-based cell-free system reveals a trade-off between transcription and translation. View ORCID ... Cell-free transcription-translation (TX-TL) systems have been used for diverse applications, from prototyping gene circuits to ... Probing metabolism in an E. coli-based cell-free system reveals a trade-off between transcription and translation ... Probing metabolism in an E. coli-based cell-free system reveals a trade-off between transcription and translation ...
Your free access has ended.. Thank you for taking us up on our offer of free access to JoVE Education until June 15th. Your ... Your JoVE Unlimited Free Trial. Fill the form to request your free trial. ... You have unlocked a 2-hour free trial now. All JoVE videos and articles can be accessed for free. ... Please follow the link in the email to activate your free trial account. If you do not see the message in your inbox, please ...
UCLA researchers in the Department of Chemistry and Biochemistry have developed a system for regulation of cofactors in cell ... Therefore, an optimized system with high yield is desired.. Innovation. UCLA researchers have developed a cell-free approach ... A Molecular Rheostat Design that Maintains ATP Levels Needed to Drive Cell-Free Synthetic Biochemistry Systems. Tech ID: 27581 ... researchers in the Department of Chemistry and Biochemistry have developed a system for regulation of cofactors in cell free ...
The incorporation of newly synthesized protein into myofibrils has been examined in a cell-free system. Myofibrils were added ... Posttranslational incorporation of contractile proteins into myofibrils in a cell-free system.. J Cell Biol 1 August 1988; 107 ... Posttranslational incorporation of contractile proteins into myofibrils in a cell-free system. M Bouché, M Bouché ... The incorporation of newly synthesized protein into myofibrils has been examined in a cell-free system. Myofibrils were added ...
We have developed a cell-free system that induces the morphological transformations characteristic of apoptosis in isolated ... Nuclear events of apoptosis in vitro in cell-free mitotic extracts: a model system for analysis of the active phase of ... We have developed a cell-free system that induces the morphological transformations characteristic of apoptosis in isolated ... Nuclear events of apoptosis in vitro in cell-free mitotic extracts: a model system for analysis of the active phase of ...
Mammalian Cell-Free System Recapitulates the Early Events of Post-Fertilization Sperm Mitophagy. , BrandiBrandi , 0 Comment ... Consequently, we anticipated that sperm mitophagy may very well be reconstituted in a cell-free system consisting of ... PREVIOUS Previous post: Mammalian Cell-Free System Recapitulates the Early Events of Post-Fertilization Sperm Mitophagy ... occasions of post-fertilization sperm mitophagy noticed in porcine zygote will be reconstituted in a cell-free system, which ...
Cell-free systems only use bacterial extract.. Mar 25, 2022 , Cell-Free Biomyths ... Cell-free systems are slower than an E.coli cell-based system ... Cell-free systems offer a wide choice of cell extracts to be ... Escherichia coli (E. Coli) is the most popular cell extract source for cell-free expression system. However, extracts from ... Cell-free eukaryotic systems for the production, engineering and modification of scFv antibody fragments. Engineering in Life ...
The MassIMO project "Cell Free Massive MIMO Systems" creates the scientific and technical foundations for the mMIMO technology ... This new, cell-free multi-antenna technology for dense interference networks can be used, for example, in an industrial ... The cell-free massive MIMO technology (mMIMO) provides high data rates, high reliability, low latencies and high energy ...
Trumpet is an operating system for simple and robust cell-free biocomputing *Judee A. Sharon ... Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing ... Khodor, J. & Gifford, D. K. Design and implementation of computational systems based on programmed mutagenesis. Biosystems 52, ... Shapiro, E. & Karunaratne, K. S. G. Method and system of computing similar to a Turing machine. US Patent 6,266,569 (2001). ...
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... meaning that red blood cells play a larger role in the immune system than previously thought. ... The study found that this DNA-binding capability triggers removal of the cells from circulation, driving inflammation and ... Red Blood Cells Play Significant Role in Immune System, Bind Cell-Free DNA. October 26, 2021. Skylar Kenney, Assistant Editor ... Red blood cells function as immune sensors by binding cell-free DNA called nucleic acid, which is present in the bodys ...
Turning your current cell phone into a feature-rich business phone system is incredibly easy. Download our free ebook to learn ... Free eBook to 4 Easy Steps to Turn Your Cell Phone Into a Virtual Phone System. ... Request Your Free eBook Now: "4 Easy Steps to Turn Your Cell Phone Into a Virtual Phone System". Turning your current cell ... Free eBook: "4 Easy Steps to Turn Your Cell Phone Into a Virtual Phone System". Turning your current cell phone into a feature- ...
... Hellmich ... Single Cell Manipulation, Analytics and Label-free Protein Detection in Microfluidic Devices for Systems NanoBiology. In N. ... "Single Cell Manipulation, Analytics and Label-free Protein Detection in Microfluidic Devices for Systems NanoBiology". In ... "Single Cell Manipulation, Analytics and Label-free Protein Detection in Microfluidic Devices for Systems NanoBiology". ...
Systems Biology, Genome-Scale Models, and Metabolic Engineering Abstract chapter 16. ,. 12 pages. Cell-Free Systems for ... Engineering Multifunctional Enzyme Systems for Optimized Metabolite Transfer between Sequential Conversion Steps Abstract ...
A novel, feeder free, autologous system for maintenance of human embryonic stem cell phenotype. European Cells and Materials, ... A novel, feeder free, autologous system for maintenance of human embryonic stem cell phenotype ...
OnePot PURE Cell-Free System. L. Grasemann; B. Lavickova; M. C. Elizondo-Cantu; S. J. Maerkl ... Bottom-Up Construction of Complex Biomolecular Systems With Cell-Free Synthetic Biology. N. Laohakunakorn; L. Grasemann; B. ... A Simple, Robust, and Low-Cost Method To Produce the PURE Cell Free System. B. Lavickova; S. J. Maerkl ... microfluidics, systems biology, cell-free synthetic biology, transcription factor biophysics / transcriptional regulation, ...
Developed on-chip cell sorting system based on an analysis of microscopic image and impact-free sorting for living cells. In ... Developed on-chip cell sorting system based on an analysis of microscopic image and impact-free sorting for living cells」の研究 ... Developed on-chip cell sorting system based on an analysis of microscopic image and impact-free sorting for living cells. / ... Developed on-chip cell sorting system based on an analysis of microscopic image and impact-free sorting for living cells. ...
A Cell-free Infection System to Study Translation, Replication and Phage-particle Production during Infection of E. coli By ... The cell-free infection systems were based on cell-free protein expression systems: two commercial systems and one custom-made ... Cell-free synthetic biology; Cell-free protein expression; Cell-free infection system; Self-assembly; RT-PCR; [14C] labelling; ... Cell-free synthetic biology articles; Cell-free protein expression articles; Cell-free infection system articles; Self-assembly ...
... such as those that regulate the eukaryotic cell division cycle. We describe here one modeling approach based on expressing the ... mathematical modeling can provide a powerful approach for investigating complex cell signaling networks, ... Cell Cycle / physiology* * Cell-Free System / chemistry * Cell-Free System / metabolism * Computer Simulation* ... Mathematical modeling as a tool for investigating cell cycle control networks Methods. 2007 Feb;41(2):238-47. doi: 10.1016/j. ...
2D TCR-pMHC dissociation kinetics of peptides of various activating potency in a cell-free system in the force range (6 to 15 ... jats:p,The T cell receptor (TCR)-peptide-MHC (pMHC) interaction is the only antigen-specific interaction during T lymphocyte ... TCR-pMHC kinetics under force in a cell-free system show no intrinsic catch bond, but a minimal encounter duration before ... TCR-pMHC kinetics under force in a cell-free system show no intrinsic catch bond, but a minimal encounter duration before ...
... cell-free system to create biosynthetic pathways to build and study sugar structures. ... cell-free system to create biosynthetic pathways to build and study sugar structures. ... cell-free system to create biosynthetic pathways to build and study sugar structures. ... cell-free system to build and study these pathways. Called GlycoPRIME, the system could lead to faster development of ...
It presents a generic model of animal cell with organelles descriptions. The size of the cell is related... ... Download The Cell for iOS to the Cell is a 3D scale model of a cell. ... Operating Systems. Operating Systems iOS. Additional Requirements Compatible with iPhone 3Gs, iPhone 4. iTunes account required ... The Cell is a 3D scale model of a cell. It presents a generic model of animal cell with organelles descriptions. The size of ...
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Opportunistic AP Selection in Cell-Free Massive MIMO-OFDM Systems In: 2022 IEEE 95th Vehicular Technology Conference. IEEE ... In: IEEE Transactions on Cognitive and Developmental Systems (TCDS), Vol. 14, No. 2, Pages 255-257, IEEE, 6/2022. ... In: 5th IEEE International Conference on Industrial Cyber-Physical Systems. IEEE International Conference on Industrial Cyber- ... Physical Systems (ICPS-2022), 5th IEEE International Conference on Industrial Cyber-Physical Systems, May 24-26, Warwick in ...
Cell-Free Protein Synthesis System Protocol. Achieve high protein yields with the ALiCE® Cell-Free Protein Synthesis System ... Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes. A video about how ... fluorescent cell tracking dyes in combination with flow and image cytometry to study interactions and fates of different cell ...
8th_of_7 : ,, the immune system can sense cell- free RNA fragments..... ... the immune system can sense cell- free RNA fragments.. -. sonam, Apr 03 2020. ... either the immune system squashes it, or it doesnt No, because Im sure the immune system needs all the leg up it can get. ... Metabolism is not free. By reducing viral load in important areas (throat and nose), we get a system better prepared to fight ...
... expression systems have been valuable tools for understanding how transcription/translation can be regulated in living cells. ... cell-free gene expression systems are one of the most widely used techniques in molecular biology1,2,3. The efficiency of cell- ... Marshall, R. & Noireaux, V. Quantitative modeling of transcription and translation of an all-E. coli cell-free system. Sci. Rep ... Cell-free gene expression systems have been valuable tools for understanding how transcription/translation can be regulated in ...
  • 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)
  • Department of Cell Biology and Anatomy, Cornell University Medical College, New York 10021. (rupress.org)
  • In: Microfluidic Applications in Biology: From Technologies to Systems Biology. (uni-bielefeld.de)
  • Single Cell Manipulation, Analytics and Label-free Protein Detection in Microfluidic Devices for Systems NanoBiology" in Microfluidic Applications in Biology: From Technologies to Systems Biology , Lion, N., Roussier, J. S., and Girault, H. eds. (uni-bielefeld.de)
  • His lab is currently working at the interface of micro-engineering, systems biology, synthetic biology, and molecular diagnostics. (epfl.ch)
  • Milan Mrksich , the Henry Wade Rogers Professor of Biomedical Engineering, Chemistry, and Cell and Molecular Biology at Northwestern's McCormick School of Engineering, is a co-author on the paper. (northwestern.edu)
  • Matthew is working on a joint research collaboration with Richard Murray's lab at Caltech and funded by the NSF and EPSRC, the project is focused on developing new metrology for synthetic biology cell free systems. (imperial.ac.uk)
  • During his PhD at the University of Sussex (2011-2016), Bilge employed genetics and cell biology to study meiosis. (imperial.ac.uk)
  • Dr. Nguyen (pronounced as "Gwen") earned a bachelor degree in Biochemistry and Cell Biology at the University of California, San Diego, and a PhD in Integrated Biology and Disease at the University of Southern California, Los Angeles. (selectbiosciences.com)
  • The QIAseq Library Quant System is intended for molecular biology applications. (qiagen.com)
  • A cell-free system is an in vitro tool widely used to study biological reactions that happen within cells apart from a full cell system, thus reducing the complex interactions typically found when working in a whole cell. (wikipedia.org)
  • Nuclear events of apoptosis in vitro in cell-free mitotic extracts: a model system for analysis of the active phase of apoptosis. (rupress.org)
  • Aglycosylated antibodies and antibody fragments produced in a scalable in vitro transcription-translation system. (synthelis.com)
  • After her graduate studies, she joined Prof Karl Tryggvason's laboratory at Duke-NUS Medical School as a research fellow in 2013 where her main focus has been to culture endothelial cells for therapeutic purposes and developing different cell models for in vitro drug testing. (selectbiosciences.com)
  • E. coli, P. pastoris , Mammalian cells, Plant cells and in vitro by cell free synthesis. (lu.se)
  • where a cell-free translation system based on Escherichia coli (E. coli), of the cell extract-based type, had the mRNA template degrade very quickly and led to the halt of protein synthesis. (wikipedia.org)
  • Additionally, cell-free protein synthesis is becoming a new alternative choice for fast protein synthesis. (wikipedia.org)
  • Under translation inhibition, transcriptional output was uniform across Mg +2 and 3-PGA concentrations, but in a translation-only system, maximum protein production occurred in the previously found optimal regime of Mg +2 and 3-PGA, suggesting a TX-TL trade-off. (biorxiv.org)
  • RMM has a financial stake in Tierra Biosciences, a private company that makes use of cell-free technologies such as those described in this article for protein expression and screening. (biorxiv.org)
  • The incorporation of newly synthesized protein into myofibrils has been examined in a cell-free system. (rupress.org)
  • Out of those 21 proteins, the interplay of seven host proteins with DEV gC protein was validated utilizing membrane-bound split- ubiquitin yeast two-hybrid system and bimolecular fluorescence complementation (BiFC) analyses. (eqalix.com)
  • Other mammalian cell lines are also more recently used for cell-free protein biosynthesis. (synthelis.com)
  • Cell-free systems offer a wide choice of cell extracts to be determined based on protein properties and associated constraints. (synthelis.com)
  • Since they present different properties and advantages, the source of cell extract will be selected according to the properties of the protein to synthesize, such as the need for post- translational modifications, and other constraints, like yield, costs or convenience of cell extract preparation (Carlson et al. (synthelis.com)
  • Firstly, the origin and biochemical characteristics of the protein to synthesize is decisive in the choice of the cell extract source (Table 1). (synthelis.com)
  • In addition, cell extracts from E. coli and wheat germ achieve the highest protein yields, well above those obtained with other eukaryotic cell-based systems. (synthelis.com)
  • In summary, the cell extract source has an important impact on protein biosynthesis, in particular for the expression feasibility, yield and cost. (synthelis.com)
  • Thus, the protein properties and production constraints should be carefully analyzed ahead of the selection of the cell extract source to be tested in cell-free expression system experiments. (synthelis.com)
  • Cell-free protein synthesis: Applications come of age. (synthelis.com)
  • Expression without boundaries: cell- free protein synthesis in pharmaceutical research. (synthelis.com)
  • Cell-free protein synthesis: pros and cons of prokaryotic and eukaryotic systems. (synthelis.com)
  • Hellmich W, Pelargus C, Leffhalm K, Ros A, Anselmetti D. Single Cell Manipulation, Analytics and Label-free Protein Detection in Microfluidic Devices for Systems NanoBiology. (uni-bielefeld.de)
  • The cell-free infection systems were based on cell-free protein expression systems: two commercial systems and one custom-made system. (fortunejournals.com)
  • The researchers used this process to develop a protein vaccine candidate modified with a sugar structure that could trigger the immune system, as well as a therapeutic antibody fragment with a sugar that can stabilize proteins as they circulate in the body. (northwestern.edu)
  • In the present study, we evaluated the efficiency of protein translation, focusing on the effect of DNA templates of different lengths on cell-free protein synthesis using a luciferase assay. (nature.com)
  • Researchers have found the M2 protein, a signature feature of influenza, interferes with the lung cells' function which inhibits the lung's ability to remove fluid. (constantcontact.com)
  • Here, we address this challenge by establishing a bacterial cell-free protein synthesis platform that enables rapid production of a variety of OSTs in their active conformations. (biorxiv.org)
  • And you instruct the cells to make the protein or proteins that you need. (medscape.com)
  • rsos from in silico dynamical modelling by tuning parameters for protein concentrations and other factors .r o y involved in the rate equations describing the systems. (lu.se)
  • Lund Protein Production Platform (LP3) has recently published its standard operating procedures for recombinant protein expression using the Baculovirus Expression Vector System (BEVS) in insect cells. (lu.se)
  • Notably, in work leading to a Nobel prize the Nirenberg and Matthaei experiment used a cell-free system, of the cell extract-based type, to incorporate chosen amino acids tagged radioactively into synthesized proteins with 30S extracted from E. coli. (wikipedia.org)
  • with prokaryotic and eukaryotic version of their cell-free translation system, have also synthesized proteins with increased production, incorporating techniques like continuous flow to add materials and remove products. (wikipedia.org)
  • With such advances in yield, productivity applications have been expanded, such as the synthesis of fusion proteins to potentially serve as vaccines for B-cell lymphomas. (wikipedia.org)
  • Posttranslational incorporation of contractile proteins into myofibrils in a cell-free system. (rupress.org)
  • For example, proteins containing disulfide bonds can be expressed in all systems, except in rabbit reticulocytes. (synthelis.com)
  • In contrast, proteins requiring post-translational modifications such as glycosylation have to be expressed in cell extracts from rabbit reticulocytes, insect cells or CHO cells. (synthelis.com)
  • Although the potential to produce complex proteins is increased using eukaryotic cell-free systems, Escherichia coli is the cell type of bacteria which presents many advantages as a source of cell extract. (synthelis.com)
  • We studied the course of infection by analysing the production of viral RNA, proteins and phage particles produced in the cell-free reactions. (fortunejournals.com)
  • Specifically, the viral RNA was replicated, the viral proteins were translated, and infectious phage particles were produced in the cell-free infection systems. (fortunejournals.com)
  • Virus particles have a nucleic acid genome and may also carry a few viral proteins, which are enclosed in a capsid of viral proteins or an envelope comprised of a small part of the host cell membrane. (fortunejournals.com)
  • Sugar structures allow these proteins to remain stable while enabling them to perform tasks, like attack a cancer cell or retrain the immune system. (northwestern.edu)
  • These mammalian cells naturally produce glycosylated proteins, but are slow-growing and can be difficult to engineer, limiting the number and diversity of glycosylation structures that can be built and tested. (northwestern.edu)
  • Jewett's lab has developed cell-free systems that create enzymes needed to create certain proteins, but up until now, these processes could not create glycosylated products without the need to reengineer living cells. (northwestern.edu)
  • In just a few months, Kightlinger used the system to construct 37 pathways, creating 23 unique sugar structures, 18 of which have never been synthesized on proteins. (northwestern.edu)
  • Gene expression in living cells is strictly self-regulated to ensure that the correct amounts of proteins are made at the most appropriate timing and location for maintaining cellular homeostasis. (nature.com)
  • He is working on developing a workflow for prototyping proteins using cell-free systems and machine learning. (imperial.ac.uk)
  • Furthermore, we demonstrate the ability of cell-free synthesized OSTs to glycosylate multiple target proteins with varying N -glycosylation acceptor sequons. (biorxiv.org)
  • Proteins targeted to the peroxisome (in plants called glyoxisomes) are synthesized in the cytoplasm of the cell and are targeted to the peroxisome post-translationally, possibly via multiple pathways. (tcdb.org)
  • Clinical manifestations of PNH occur when a HSC clone carrying somatic PIGA mutations acquires a growth advantage and differentiates, generating mature blood cells that are deficient of GPI-anchored proteins. (medscape.com)
  • E. coli, wheat germ, and rabbit reticulocytes have all proven useful to create cell-free systems by extraction of their interior components. (wikipedia.org)
  • Motivated by a minimal model of cell-free metabolism, this work explored the effects of energy molecules, which power TX-TL, and fuel molecules, which regenerate energy by harnessing core metabolism, on an E. coli -based TX-TL system. (biorxiv.org)
  • Escherichia coli (E. Coli) is the most popular cell extract source for cell-free expression system. (synthelis.com)
  • Eukaryotic and prokaryotic cell internals have been used for creation of these simplified environments. (wikipedia.org)
  • As an illustration, the production of full-length and smaller formats of antibodies exploits the advantages of either prokaryotic or eukaryotic cell-free expression systems (Stech et al. (synthelis.com)
  • Cell-free eukaryotic systems for the production, engineering and modification of scFv antibody fragments. (synthelis.com)
  • Although not a traditional experimental "method," mathematical modeling can provide a powerful approach for investigating complex cell signaling networks, such as those that regulate the eukaryotic cell division cycle. (nih.gov)
  • Similarly, wheat germ has been ground with acid-washed sand or powdered glass to open the cell membranes up. (wikipedia.org)
  • However, extracts from wheat germ, rabbit reticulocytes or insect cells are also commonly used. (synthelis.com)
  • UCLA researchers in the Department of Chemistry and Biochemistry have developed a system for regulation of cofactors in cell free biochemical production. (universityofcalifornia.edu)
  • This system may provide a powerful means of dissecting the biochemical mechanisms underlying the final stages of apoptosis. (rupress.org)
  • While this relationship is often true in physics models, a . quantitative relationship between the biochemical dynamics and the free energy landscape has not open been widely exploited in developmental processes. (lu.se)
  • They rely on their target cell for molecular building blocks, energy sources and host factors to replicate, since viruses are gene poor compared to their host [1]. (fortunejournals.com)
  • This review is intended to justify the place of mathematical modeling as a standard method for studying molecular regulatory networks and to guide the non-expert to initiate modeling projects in order to gain a systems-level perspective for complex control systems. (nih.gov)
  • Monisha was a Marie Curie Sklodowska Fellow within the ITN Translocation project and was enrolled in a PhD program at the Institute for Cell and Molecular Biosciences (ICaMB), Newcastle University, UK. (imperial.ac.uk)
  • That is, life is an emergent property of some molecular systems characterized by a very peculiar type of structural and dynamical (self-)organization. (frontiersin.org)
  • The design exploits the unusual electrical properties of structures called single-wall carbon nanotubes, using them as "molecular wires in light harvesting cells," said Choi, whose research group is based at the Birck Nanotechnology and Bindley Bioscience centers at Purdue's Discovery Park. (sciencedaily.com)
  • Two elements are critical for the technology to mimic nature's self-repair mechanism: molecular recognition and thermodynamic metastability, or the ability of the system to continuously be dissolved and reassembled. (sciencedaily.com)
  • Caoimhe's project is focused on developing metrology standards for mammalian cell-free systems. (imperial.ac.uk)
  • Killing viral particles, as well as cells infected by the virus, in the throat, tonsils and trachea would lower the viral load the immune system must fight. (halfbakery.com)
  • Frances Sheridan Goulart author of "Super Immunity Foods: A Complete Program to Boost Wellness, Speed Recovery and Keep Your Body Strong," notes that one of the most beneficial aspects of eating berries is that they offer a whole host of antioxidants that can boost your immune system. (constantcontact.com)
  • It is needed by the body to support the immune system and for blood clotting. (medlineplus.gov)
  • And then we make a product just for you to educate your immune system, your T cells, to recognize the mutation of a cancer cell that they have missed so far, so that your immune system can go to work and get rid of your cancer. (medscape.com)
  • These issues stem largely from the fact that cell lysate contains an active and complex metabolism whose effect on TX-TL has remained largely uncharacterized. (biorxiv.org)
  • Reprogramming is done with feeder free system. (lu.se)
  • The skin manifestations occur along the lines of Blaschko, which represent the routes of embryonic cell migration. (medscape.com)
  • Recently, a book chapter with protocols for the Baculovirus Expression Vector System in insect cells was published by LP3. (lu.se)
  • Cell-free systems only use bacterial extract. (synthelis.com)
  • RNeasy Protect Mini Kits enable stabilization of RNA in tissue samples, RNA and DNA in sorted or cultured cells, RNA in human saliva samples and RNA in bacterial samples. (qiagen.com)
  • The array format provides this standard in five predispensed, sequential 10-fold dilutions mixed with a PCR primer assay in triplicate, ensuring that your sample library will fall within the detection range of the array (see figure Principle of the QIAseq Library Quant System The serial dilutions of the DNA standard (5 sequential 10-fold dilutions) generate a standard curve. The sample library should fall within the standard curve. "> Principle of QIAseq Library Quant System ). (qiagen.com)
  • Further, Calhoun and Swartz were able to use a glycolytic intermediate to fuel a cell-free system, enabling relatively inexpensive ATP generation compared to reagent usage in phosphoenolpyruvate reactions. (wikipedia.org)
  • Reactions are typically assembled in nuclease-free 1.5 mL microcentrifuge tubes. (neb.com)
  • In the last decade, the chemists have been delighted by the catalytic orchestration found in vivo, and have isolated multi-enzyme system to work ex-vivo in both natural and non-natural tandem reactions creating a new concept: systems biocatalysis. (ibecbarcelona.eu)
  • We have paid our attention to the heterogenization of multi-enzyme system to catalyze tandem reactions. (ibecbarcelona.eu)
  • Cell-free biosystems have several advantages suitable in industrial applications: Very high product yields are usually accomplished without the formation of by-products or the synthesis of cell mass. (wikipedia.org)
  • Cell-Free Synthesis Meets Antibody Production: A Review. (synthelis.com)
  • The system uses extracts prepared from mitotic chicken hepatoma cells following a sequential S phase/M phase synchronization. (rupress.org)
  • We illustrate this approach throughout with the simple and well-characterized example of mitotic cell cycles in frog egg extracts. (nih.gov)
  • Although Dr. Goulart mentions wild blueberries as having one of the highest levels of antioxidants, it is the Aroniaberry that checks in with a much higher level of antioxidants and a more balanced protection against free radicals that can limit our bodies ability to fight illness. (constantcontact.com)
  • It targets human's lung cells through a system that involves free radicals. (constantcontact.com)
  • By eliminating these free radicals, the flu virus weakens and cannot compromise the cells of the lungs. (constantcontact.com)
  • It works as an antioxidant to protect your cells against free radicals. (medlineplus.gov)
  • UCLA researchers have developed a cell-free approach for production of chemicals. (universityofcalifornia.edu)
  • But because researchers have used mammalian cells to create the biosynthetic pathways (sets of enzyme catalysts) to build and study these sugar structures, the process has historically taken a long time and has required specialized laboratory equipment. (northwestern.edu)
  • Northwestern Engineering researchers have now developed a quick, cell-free system to build and study these pathways. (northwestern.edu)
  • Researchers who have spent the last two years studying the security of car computer systems have revealed that they can take control of vehicles wirelessly. (technologyreview.com)
  • Researchers have previously shown they can take control of a car's dashboard display, among other systems. (technologyreview.com)
  • The researchers were able to control everything from the car's brakes to its door locks to its computerized dashboard displays by accessing the onboard computer through GM's OnStar and Ford's Sync, as well as through the Bluetooth connections intended for making hands-free phone calls. (technologyreview.com)
  • With the new work, the researchers systematically analyzed ways they could get at a car's computer systems without having physical access. (technologyreview.com)
  • The researchers attacked the car's Bluetooth system, which allows a driver to make hands-free cell-phone calls. (technologyreview.com)
  • The researchers found that they could take control of this system by breaking through its authentication system. (technologyreview.com)
  • We were surprised to find that the attack surface was so broad," Kohno says, referring to the wide variety of ways the researchers were able to gain access to the car's computer systems. (technologyreview.com)
  • Louis Lanzerotti , a distinguished research professor in the physics department at the New Jersey Institute of Technology and the chair of the Committee on Electronic Vehicle Controls and Unintended Acceleration, says the researchers were invited to speak at the event as part of its review of electronic vehicle controls, systems, and safety across the industry. (technologyreview.com)
  • Attacking cars remotely significantly broadens the threat and greatly increases the impact of this and [the researchers'] previous work," says Aurélien Francillon , a researcher in the system security group at ETH Zurich, in Switzerland, who has also worked on automotive security. (technologyreview.com)
  • Researchers are creating a new type of solar cell designed to self-repair like natural photosynthetic systems in plants by using carbon nanotubes and DNA, an approach aimed at increasing service life and reducing cost. (sciencedaily.com)
  • Enzymatic systems, without the barrier of cellular membrane, usually have faster reaction rates than microbial systems. (wikipedia.org)
  • To elucidate the complex mechanism of gene regulation in living cellular systems, it is necessary to closely investigate each phase of gene expression and also to shed light on the cooperative effects between transcription and translation. (nature.com)
  • Data from this study indicate that it is possible to predict acute in vivo inflammatory potential of NP with cell-free and cellular assays by using NP surface area-based dose and response metrics, but that a cellular component is required to achieve a higher degree of predictive power. (cdc.gov)
  • Nobel prize winner Eduard Buchner was arguably the first to present a cell-free system using yeast extracts, but since then alternative sources have been found. (wikipedia.org)
  • Consequently, we anti cipated that sperm mitophagy may very well be reconstituted in a cell-free system consisting of permeabilized mammalian spermatozoa co-incubated with porcine oocyte extracts. (eqalix.com)
  • Extracellular vesicles (EVs), including exosomes, are membranous particles released by cells into the extracellular space. (mdpi.com)
  • In this study, we have analysed the production of infectious Qβ phage particles in three different cell-free infection systems upon addition of the Qβ genome as a template. (fortunejournals.com)
  • Infectious Qβ phage particles were produced at yields of 2.5 × 105 PFU/μL reaction and 2.5 × 103 PFU/μL reaction in the commercial and custom-made system, respectively. (fortunejournals.com)
  • The cell-based systems also used electron spin resonance (ESR) as well as luciferase reporter activity to rank the different particle types in comparison to benchmark particles of low and high activity. (cdc.gov)
  • This work represents a significant advancement in understanding the effects of fuel and energy metabolism on TX-TL in cell-free systems and lays the foundation for improving TX-TL performance, lifetime, standardization, and prediction. (biorxiv.org)
  • Two cell-free and two cell-based assays were evaluated for their power in predicting in vivo toxicity of eight distinct particle types with widely differing physicochemical characteristics. (cdc.gov)
  • Engineering of metabolic processes have been achieved through cell-free systems. (wikipedia.org)
  • Type 1 diabetes mellitus is a metabolic disease resulting from the destruction of insulinproducing β cells in the pancreas, that leads to hyperglycemia [1,2,20]. (researchgate.net)
  • The RNeasy Protect Mini, RNeasy Protect Cell Mini and RNeasy Protect Bacteria Mini Kits can be automated on the QIAcube Connect . (qiagen.com)
  • Autopoiesis thus becomes a convenient and elegant theoretical framework to guide the variegate experimental efforts to fabricate synthetic (artificial) cells (SCs/ACs), especially with respect to studies oriented at the origins and emergence of life. (frontiersin.org)
  • This article discusses what was formerly referred to as incontinentia pigmenti type 2, also known as Bloch-Sulzberger syndrome, a rare, X-linked, dominantly inherited disorder of skin pigmentation that is often associated with ocular, dental, and central nervous system abnormalities. (medscape.com)
  • Cell-free gene expression systems have been valuable tools for understanding how transcription/translation can be regulated in living cells. (nature.com)
  • Cell-free systems may be divided into two primary classifications: cell extract-based, which remove components from within a whole cell for external use, and purified enzyme-based, which use purified components of the molecules known to be involved in a given process. (wikipedia.org)
  • These outcomes reveal that the early developmental occasions of post-fertilization sperm mitophagy noticed in porcine zygote will be reconstituted in a cell-free system, which may grow to be a useful gizmo for figuring out extra molecules that regulate mitochondrial inheritance in mammals. (eqalix.com)
  • The cells contain light-absorbing dyes called chromophores, chlorophyll-like molecules that degrade due to exposure to sunlight. (sciencedaily.com)
  • The QIAseq Library Quant System uses real-time PCR to specifically quantify DNA molecules with sequencing adaptors at both ends, ensuring optimal cluster density or template-to-bead ratio. (qiagen.com)
  • The QIAseq Library Quant System, by contrast, uses real-time PCR to quantify only DNA molecules with adaptors at both ends, which are the only amplifiable molecules during emulsion PCR (Ion Torrent platform) or bridge PCR (Illumina platform), and therefore provides highly accurate quantification of amplifiable library molecules. (qiagen.com)
  • Programming cells by multiplex genome engineering and accelerated evolution. (nih.gov)
  • Handling, maintenance and understanding of human pluripotent stem cells. (lu.se)
  • Paroxysmal nocturnal hemoglobinuria (PNH) is a clonal hematopoietic stem cell (HSC) disease. (medscape.com)
  • Allogeneic hematopoietic stem cell transplantation is curative but it requires a histocompatible donor and is associated with significant morbidity and mortality, so it is reserved for severe cases of PNH with aplastic anemia or transformation to leukemia. (medscape.com)
  • those of cell reprogramming, thereby avoiding exhaustive trial- energy landscape, deterministic models, and-error simulations with rate equations for different stem cell commitment, reprogramming parameter sets. (lu.se)
  • We explore the method on three circuits for haematopoiesis and embryonic stem cell development for commitment and reprogramming scenarios and illustrate how the method can be used to determine sequential steps for onsets of external factors, essential for efficient reprogramming. (lu.se)
  • Several (unsuccessful) attempts to build cell-like systems of minimal complexity fill the annals of science ( Hanczyc, 2009 ). (frontiersin.org)
  • The sensitivity and dynamic range of the QIAseq Library Quant System were compared against Agilent's High Sensitivity DNA Kit, using two NGS libraries of differing concentrations. (qiagen.com)
  • By contrast, the QIAseq Library Quant Array was able to quantify both libraries (see figure The QIAseq Library Quant System enables quantification of libraries with concentrations below the detection limit of conventional methods The QIAseq Library Quant Array's high sensitivity and broad dynamic range enable the quantification of both the NGS-L1 and NGS-L2 libraries [A]. By contrast, Agilent's High Sensitivity DNA Kit (for use with the Agilent 2100 BioAnalyzer) quantified only the NGS-L1 library; with this kit, the NGS-L2 concentration was too low for quantification [B]. "> QIAseq Library Quant System enables quantification of libraries with concentrations below the detection limit of conventional methods ). (qiagen.com)
  • The system could also be used to provide modular, on-demand biomanufacturing platforms that provide medicines or vaccines in resource-limited settings. (northwestern.edu)
  • Discussion also centred on the role of inactivated poliovirus vaccines in the eradication program and the maintenance of a poliovirus-free world, whenever this goal should be achieved. (springer.com)
  • Cell-free synthetic pathway biotransformation biosystems are proposed as a new low-cost biomanufacturing platform compared to microbial fermentation used for thousands of years. (wikipedia.org)
  • degradation is considered mediated by the interaction between the ubiquitin -proteasome system (UPS) and the autophagic pathway. (eqalix.com)
  • We used a laminar flow chamber to measure, first, 2D TCR-pMHC dissociation kinetics of peptides of various activating potency in a cell-free system in the force range (6 to 15 pN) previously associated with catch-slip transitions and, second, 2D TCR-pMHC association kinetics, for which the method is well suited. (ox.ac.uk)
  • Co-immobilization of multi-enzyme systems improve: 1) the kinetics of the chemical cascades due to the spatial localization of the different biocatalytic modules that avoids intermediate accumulation and increases cofactor recycling efficiency, 2) the stability of the biocatalysts due to both structural rigidification and in situ elimination of toxic by-products, 3) the biocatalyst recycle and 4) the biocatalyst adaptation to continuous processes. (ibecbarcelona.eu)
  • Michaelis-Menten or Hill kinetics do not have a corresponding free energy from which the rate equations i. (lu.se)
  • The optimal design of the immobilization protocols enables co-immobilizing several enzymes and cofactors on the porous carrier to optimize their spatial localization across the carrier microstructure and preserve both global activity and stability of the multi-enzyme systems. (ibecbarcelona.eu)
  • We have developed a cell-free system that induces the morphological transformations characteristic of apoptosis in isolated nuclei. (rupress.org)
  • Both processes are inhibited by Zn2+, an inhibitor of apoptosis in intact cells. (rupress.org)
  • Nuclear lamina disassembly accompanies these structural changes in added nuclei, and we show that lamina disassembly is a characteristic feature of apoptosis in intact cells of mouse, human and chicken. (rupress.org)
  • [ 6 ] NF-KB protects cells from apoptosis in response to tumor necrosis factor-alpha (TNF-alpha). (medscape.com)
  • In the skin, NF-KB regulates cell growth in the stratified epithelium and apoptosis. (medscape.com)
  • The invited scientists reviewed the pathfinding role of PV in virology, evaluated the progress made in eradicating this devastating human pathogen and discussed strategies to achieve the goal of a polio-free world. (springer.com)
  • To a closer inspection, in fact, the Authors put forward an operational methodology for the construction of chemical reacting systems that would show the difficult-to-define property of being alive just by fulfilling a specific structural and dynamic organization. (frontiersin.org)
  • On-Demand Webinar on Serum-free Expansion of NK Cells with K562- Based Feeder Systems for Clinical Applications featuring Dean Lee, MD, Nationwide Children's Hospital. (thermofisher.com)
  • However, the low yields and productivity, in part due to the complexity of living systems has limited their industrial applications. (universityofcalifornia.edu)
  • 2015). High production yields have been obtained using prokaryotic systems in the presence of PDI or DsbC and variable ratios of GSSG and GSH: up to 950 mg/L for a scFv, 300 mg/L for a Fab and 400 mg/L of an aglycosylated IgG were obtained (Yin et al. (synthelis.com)
  • 2012). In addition, the production of functional scFv antibody fragments has been achieved by using vesicle- containing Spodoptera frugiperda (Sf21) insect cell-free expression systems but with yields of 10-15 μg/mL approximately (Stech et al. (synthelis.com)
  • Numbers in parentheses denote the stated expression levels of PrP c into the animal species and cell lines used. (cdc.gov)
  • Haptoglobin is produced by the liver and collects the hemoglobin from destroyed red blood cells, then transporting it back to the liver, where heme is converted to bilirubin. (medscape.com)
  • melanin collects in the dermis as free pigment or aggregates of melanophages. (medscape.com)
  • In systems where additional energy is supplied and where a fuel source is absent, the trade-off is absent, suggesting the trade-off arises from limitations in the regulation of translation and efficient energy regeneration. (biorxiv.org)
  • The preparation of cell extract is simple, robust and inexpensive. (synthelis.com)
  • Cell-free transcription-translation (TX-TL) systems have been used for diverse applications, from prototyping gene circuits to providing a platform for the development of synthetic life, but their performance is limited by issues such as batch-to-batch variability, poor predictability, and limited lifetime. (biorxiv.org)
  • Olariu V, Manesso E, commitment and Peterson C. 2017 A deterministic method for estimating free energy genetic network reprogramming paths landscapes with applications to cell commitment and reprogramming paths. (lu.se)