Genetic Complementation Test: A test used to determine whether or not complementation (compensation in the form of dominance) will occur in a cell with a given mutant phenotype when another mutant genome, encoding the same mutant phenotype, is introduced into that cell.Mutation: Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations.Genes, Bacterial: The functional hereditary units of BACTERIA.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.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.Cloning, Molecular: The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.Phenotype: The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment.Chromosome Mapping: Any method used for determining the location of and relative distances between genes on a chromosome.Plasmids: Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS.Amino Acid Sequence: The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.Genes: A category of nucleic acid sequences that function as units of heredity and which code for the basic instructions for the development, reproduction, and maintenance of organisms.Alleles: Variant forms of the same gene, occupying the same locus on homologous CHROMOSOMES, and governing the variants in production of the same gene product.Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.Bacterial Proteins: Proteins found in any species of bacterium.Mutagenesis, Insertional: Mutagenesis where the mutation is caused by the introduction of foreign DNA sequences into a gene or extragenic sequence. This may occur spontaneously in vivo or be experimentally induced in vivo or in vitro. Proviral DNA insertions into or adjacent to a cellular proto-oncogene can interrupt GENETIC TRANSLATION of the coding sequences or interfere with recognition of regulatory elements and cause unregulated expression of the proto-oncogene resulting in tumor formation.Crosses, Genetic: Deliberate breeding of two different individuals that results in offspring that carry part of the genetic material of each parent. The parent organisms must be genetically compatible and may be from different varieties or closely related species.Operon: In bacteria, a group of metabolically related genes, with a common promoter, whose transcription into a single polycistronic MESSENGER RNA is under the control of an OPERATOR REGION.Restriction Mapping: Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.DNA Transposable Elements: Discrete segments of DNA which can excise and reintegrate to another site in the genome. Most are inactive, i.e., have not been found to exist outside the integrated state. DNA transposable elements include bacterial IS (insertion sequence) elements, Tn elements, the maize controlling elements Ac and Ds, Drosophila P, gypsy, and pogo elements, the human Tigger elements and the Tc and mariner elements which are found throughout the animal kingdom.Saccharomyces cerevisiae: A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement.DNA, Bacterial: Deoxyribonucleic acid that makes up the genetic material of bacteria.Genes, Regulator: Genes which regulate or circumscribe the activity of other genes; specifically, genes which code for PROTEINS or RNAs which have GENE EXPRESSION REGULATION functions.Genes, Lethal: Genes whose loss of function or gain of function MUTATION leads to the death of the carrier prior to maturity. They may be essential genes (GENES, ESSENTIAL) required for viability, or genes which cause a block of function of an essential gene at a time when the essential gene function is required for viability.Recombination, Genetic: Production of new arrangements of DNA by various mechanisms such as assortment and segregation, CROSSING OVER; GENE CONVERSION; GENETIC TRANSFORMATION; GENETIC CONJUGATION; GENETIC TRANSDUCTION; or mixed infection of viruses.Temperature: The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms.Arabidopsis: A plant genus of the family BRASSICACEAE that contains ARABIDOPSIS PROTEINS and MADS DOMAIN PROTEINS. The species A. thaliana is used for experiments in classical plant genetics as well as molecular genetic studies in plant physiology, biochemistry, and development.Coliphages: Viruses whose host is Escherichia coli.Suppression, Genetic: Mutation process that restores the wild-type PHENOTYPE in an organism possessing a mutationally altered GENOTYPE. The second "suppressor" mutation may be on a different gene, on the same gene but located at a distance from the site of the primary mutation, or in extrachromosomal genes (EXTRACHROMOSOMAL INHERITANCE).Genes, Plant: The functional hereditary units of PLANTS.Transduction, Genetic: The transfer of bacterial DNA by phages from an infected bacterium to another bacterium. This also refers to the transfer of genes into eukaryotic cells by viruses. This naturally occurring process is routinely employed as a GENE TRANSFER TECHNIQUE.Drosophila melanogaster: A species of fruit fly much used in genetics because of the large size of its chromosomes.

The Saccharomyces cerevisiae ETH1 gene, an inducible homolog of exonuclease III that provides resistance to DNA-damaging agents and limits spontaneous mutagenesis. (1/10927)

The recently sequenced Saccharomyces cerevisiae genome was searched for a gene with homology to the gene encoding the major human AP endonuclease, a component of the highly conserved DNA base excision repair pathway. An open reading frame was found to encode a putative protein (34% identical to the Schizosaccharomyces pombe eth1(+) [open reading frame SPBC3D6.10] gene product) with a 347-residue segment homologous to the exonuclease III family of AP endonucleases. Synthesis of mRNA from ETH1 in wild-type cells was induced sixfold relative to that in untreated cells after exposure to the alkylating agent methyl methanesulfonate (MMS). To investigate the function of ETH1, deletions of the open reading frame were made in a wild-type strain and a strain deficient in the known yeast AP endonuclease encoded by APN1. eth1 strains were not more sensitive to killing by MMS, hydrogen peroxide, or phleomycin D1, whereas apn1 strains were approximately 3-fold more sensitive to MMS and approximately 10-fold more sensitive to hydrogen peroxide than was the wild type. Double-mutant strains (apn1 eth1) were approximately 15-fold more sensitive to MMS and approximately 2- to 3-fold more sensitive to hydrogen peroxide and phleomycin D1 than were apn1 strains. Elimination of ETH1 in apn1 strains also increased spontaneous mutation rates 9- or 31-fold compared to the wild type as determined by reversion to adenine or lysine prototrophy, respectively. Transformation of apn1 eth1 cells with an expression vector containing ETH1 reversed the hypersensitivity to MMS and limited the rate of spontaneous mutagenesis. Expression of ETH1 in a dut-1 xthA3 Escherichia coli strain demonstrated that the gene product functionally complements the missing AP endonuclease activity. Thus, in apn1 cells where the major AP endonuclease activity is missing, ETH1 offers an alternate capacity for repair of spontaneous or induced damage to DNA that is normally repaired by Apn1 protein.  (+info)

Impaired translesion synthesis in xeroderma pigmentosum variant extracts. (2/10927)

Xeroderma pigmentosum variant (XPV) cells are characterized by a cellular defect in the ability to synthesize intact daughter DNA strands on damaged templates. Molecular mechanisms that facilitate replication fork progression on damaged DNA in normal cells are not well defined. In this study, we used single-stranded plasmid molecules containing a single N-2-acetylaminofluorene (AAF) adduct to analyze translesion synthesis (TLS) catalyzed by extracts of either normal or XPV primary skin fibroblasts. In one of the substrates, the single AAF adduct was located at the 3' end of a run of three guanines that was previously shown to induce deletion of one G by a slippage mechanism. Primer extension reactions performed by normal cellular extracts from four different individuals produced the same distinct pattern of TLS, with over 80% of the products resulting from the elongation of a slipped intermediate and the remaining 20% resulting from a nonslipped intermediate. In contrast, with cellular extracts from five different XPV patients, the TLS reaction was strongly reduced, yielding only low amounts of TLS via the nonslipped intermediate. With our second substrate, in which the AAF adduct was located at the first G in the run, thus preventing slippage from occurring, we confirmed that normal extracts were able to perform TLS 10-fold more efficiently than XPV extracts. These data demonstrate unequivocally that the defect in XPV cells resides in translesion synthesis independently of the slippage process.  (+info)

Conserved residues of human XPG protein important for nuclease activity and function in nucleotide excision repair. (3/10927)

The human XPG endonuclease cuts on the 3' side of a DNA lesion during nucleotide excision repair. Mutations in XPG can lead to the disorders xeroderma pigmentosum (XP) and Cockayne syndrome. XPG shares sequence similarities in two regions with a family of structure-specific nucleases and exonucleases. To begin defining its catalytic mechanism, we changed highly conserved residues and determined the effects on the endonuclease activity of isolated XPG, its function in open complex formation and dual incision reconstituted with purified proteins, and its ability to restore cellular resistance to UV light. The substitution A792V present in two XP complementation group G (XP-G) individuals reduced but did not abolish endonuclease activity, explaining their mild clinical phenotype. Isolated XPG proteins with Asp-77 or Glu-791 substitutions did not cleave DNA. In the reconstituted repair system, alanine substitutions at these positions permitted open complex formation but were inactive for 3' cleavage, whereas D77E and E791D proteins retained considerable activity. The function of each mutant protein in the reconstituted system was mirrored by its ability to restore UV resistance to XP-G cell lines. Hydrodynamic measurements indicated that XPG exists as a monomer in high salt conditions, but immunoprecipitation of intact and truncated XPG proteins showed that XPG polypeptides can interact with each other, suggesting dimerization as an element of XPG function. The mutation results define critical residues in the catalytic center of XPG and strongly suggest that key features of the strand cleavage mechanism and active site structure are shared by members of the nuclease family.  (+info)

Disruption of the Toxoplasma gondii bradyzoite-specific gene BAG1 decreases in vivo cyst formation. (4/10927)

The bradyzoite stage of the Apicomplexan protozoan parasite Toxoplasma gondii plays a critical role in maintenance of latent infection. We reported previously the cloning of a bradyzoite-specific gene BAG1/hsp30 (previously referred to as BAG5) encoding a cytoplasmic antigen related to small heat shock proteins. We have now disrupted BAG1 in the T. gondii PLK strain by homologous recombination. H7, a cloned null mutant, and Y8, a control positive for both cat and BAG1, were chosen for further characterization. Immunofluorescence and Western blot analysis of bradyzoites with BAG1 antisera demonstrated expression of BAG1 in the Y8 and the PLK strain but no expression in H7. All three strains expressed a 116 kDa bradyzoite cyst wall antigen, a 29 kDa matrix antigen and the 65 kDa matrix reactive antigen MAG1. Mice inoculated with H7 parasites formed significantly fewer cysts than those inoculated with the Y8 and the PLK strains. H7 parasites were complemented with BAG1 using phleomycin selection. Cyst formation in vivo for the BAG1-complemented H7 parasites was similar to wild-type parasites. We therefore conclude that BAG1 is not essential for cyst formation, but facilitates formation of cysts in vivo.  (+info)

Isocitrate lyase of Ashbya gossypii--transcriptional regulation and peroxisomal localization. (5/10927)

The isocitrate lyase-encoding gene AgICL1 from the filamentous hemiascomycete Ashbya gossypii was isolated by heterologous complementation of a Saccharomyces cerevisiae icl1d mutant. The open reading frame of 1680 bp encoded a protein of 560 amino acids with a calculated molecular weight of 62584. Disruption of the AgICL1 gene led to complete loss of AgIcl1p activity and inability to grow on oleic acid as sole carbon source. Compartmentation of AgIcl1p in peroxisomes was demonstrated both by Percoll density gradient centrifugation and by immunogold labeling of ultrathin sections using specific antibodies. This fitted with the peroxisomal targeting signal AKL predicted from the C-terminal DNA sequence. Northern blot analysis with mycelium grown on different carbon sources as well as AgICL1 promoter replacement with the constitutive AgTEF promoter revealed a regulation at the transcriptional level. AgICL1 was subject to glucose repression, derepressed by glycerol, partially induced by the C2 compounds ethanol and acetate, and fully induced by soybean oil.  (+info)

Mitotic recombination in the heterochromatin of the sex chromosomes of Drosophila melanogaster. (6/10927)

The frequency of spontaneous and X-ray-induced mitotic recombination involving the Y chromosome has been studied in individuals with a marked Y chromosome arm and different XY compound chromosomes. The genotypes used include X chromosomes with different amounts of X heterochromatin and either or both arms of the Y chromosome attached to either side of the centromere. Individuals with two Y chromosomes have also been studied. The results show that the bulk of mitotic recombination takes place between homologous regions.  (+info)

Efflux-mediated aminoglycoside and macrolide resistance in Burkholderia pseudomallei. (7/10927)

Burkholderia pseudomallei, the causative agent of melioidosis, is intrinsically resistant to a wide range of antimicrobial agents including beta-lactams, aminoglycosides, macrolides, and polymyxins. We used Tn5-OT182 to mutagenize B. pseudomallei to identify the genes involved in aminoglycoside resistance. We report here on the identification of AmrAB-OprA, a multidrug efflux system in B. pseudomallei which is specific for both aminoglycoside and macrolide antibiotics. We isolated two transposon mutants, RM101 and RM102, which had 8- to 128-fold increases in their susceptibilities to the aminoglycosides streptomycin, gentamicin, neomycin, tobramycin, kanamycin, and spectinomycin. In addition, both mutants, in contrast to the parent, were susceptible to the macrolides erythromycin and clarithromycin but not to the lincosamide clindamycin. Sequencing of the DNA flanking the transposon insertions revealed a putative operon consisting of a resistance, nodulation, division-type transporter, a membrane fusion protein, an outer membrane protein, and a divergently transcribed regulatorprotein. Consistent with the presence of an efflux system, both mutants accumulated [3H] dihydro streptomycin, whereas the parent strain did not. We constructed an amr deletion strain, B. pseudomallei DD503, which was hypersusceptible to aminoglycosides and macrolides and which was used successfully in allelic exchange experiments. These results suggest that an efflux system is a major contributor to the inherent high-level aminoglycoside and macrolide resistance found in B. pseudomallei.  (+info)

Analysis of 4-phosphopantetheinylation of polyhydroxybutyrate synthase from Ralstonia eutropha: generation of beta-alanine auxotrophic Tn5 mutants and cloning of the panD gene region. (8/10927)

The postulated posttranslational modification of the polyhydroxybutyrate (PHA) synthase from Ralstonia eutropha by 4-phosphopantetheine was investigated. Four beta-alanine auxotrophic Tn5-induced mutants of R. eutropha HF39 were isolated, and two insertions were mapped in an open reading frame with strong similarity to the panD gene from Escherichia coli, encoding L-aspartate-1-decarboxylase (EC 4.1.1.15), whereas two other insertions were mapped in an open reading frame (ORF) with strong similarity to the NAD(P)+ transhydrogenase (EC 1.6.1.1) alpha 1 subunit, encoded by the pntAA gene from Escherichia coli. The panD gene was cloned by complementation of the panD mutant of R. eutropha Q20. DNA sequencing of the panD gene region (3,312 bp) revealed an ORF of 365 bp, encoding a protein with 63 and 67% amino acid sequence similarity to PanD from E. coli and Bacillus subtilis, respectively. Subcloning of only this ORF into vectors pBBR1MCS-3 and pBluescript KS- led to complementation of the panD mutants of R. eutropha and E. coli SJ16, respectively. panD-encoded L-aspartate-1-decarboxylase was further confirmed by an enzymatic assay. Upstream of panD, an ORF with strong similarity to pntAA from E. coli, encoding NAD(P)+ transhydrogenase subunit alpha 1 was found; downstream of panD, two ORFs with strong similarity to pntAB and pntB, encoding subunits alpha 2 and beta of the NAD(P)+ transhydrogenase, respectively, were identified. Thus, a hitherto undetermined organization of pan and pnt genes was found in R. eutropha. Labeling experiments using one of the R. eutropha panD mutants and [2-14C]beta-alanine provided no evidence that R. eutropha PHA synthase is covalently modified by posttranslational attachment of 4-phosphopantetheine, nor did the E. coli panD mutant exhibit detectable labeling of functional PHA synthase from R. eutropha.  (+info)

*Tim Stearns

... thesis work was notable for identifying exceptions to the genetic complementation test that were useful for defining genetic ... His Ph.D. advisor at MIT was David Botstein, and the title of his thesis was "Genetic analysis of the yeast microtubule ...

*Transheterozygote

... s are useful in the study of genetic interactions and complementation testing. A transheterozygote is a ... Transheterozygotes are useful in complementation testing, as pioneered by geneticist Edward B. Lewis. If a transheterozygote ... Yue L, Karr TL, Nathan DF, Swift H, Srinivasan S, Lindquist S (1999). "Genetic analysis of viable Hsp90 alleles reveals a ... Price JV, Savenye ED, Lum D, Breitkreutz A (1997). "Dominant enhancers of Egfr in Drosophila melanogaster: genetic links ...

*Complementation (genetics)

A complementation test (sometimes called a "cis-trans" test) can be used to test whether the mutations in two strains are in ... Heterosis appears to be largely due to genetic complementation, that is the masking of deleterious recessive alleles in hybrid ... The complementation test was one of the main tools used in the early Neurospora work, because it was easy to do, and allowed ... Complementation will not occur if the mutations are in the same gene. The convenience and essence of this test is that the ...

*GRE Biochemistry, Cell and Molecular Biology Test

... transduction and conjugation Recombination and complementation Mutational analysis Genetic mapping and linkage analysis B. ... GRE Biology Test GRE Chemistry Test GRE Literature in English Test GRE Mathematics Test GRE Physics Test GRE Psychology Test ... A sampling of test item content is given below: A Chemical and Physical Foundations Thermodynamics and kinetics Redox states ... Scores are scaled and then reported as a number between 200 and 990; however, in recent versions of the test, the maximum and ...

*Outline of biology

... genetic screen - DNA paternity testing - linkage map - genetic map List of biologists List of biochemists List of ecologists ... genetic mosaic - maternal effect - penetrance - complementation - suppression - epistasis - genetic linkage chromosomal effects ... genetic structure: DNA - DNA replication - nucleosome - genetic code - codon - transcription factor - transcription - ... blood test Outline of cell biology Cell biology: the cell coined by Robert Hooke Techniques: cell culture - microscope - SEM - ...

*Panellus stipticus

Using techniques of genetic complementation, Macrae paired nonluminescent monocaryons with luminescent ones, and concluded that ... After intercontinental compatibility tests, Petersen and Bermudes suggested that bioluminescence and compatibility were ... Lingle ML, Porter D, O'Kane DJ (1992). "Preliminary analysis of genetic complementation of bioluminescence in Panellus ... Genetic analysis has shown that luminescence is controlled by a single dominant allele. The luminescent glow of this and other ...

*Zellweger syndrome

In addition to genetic tests involving the sequencing of PEX genes, biochemical tests have proven highly effective for the ... A study using complementation analysis". Journal of Clinical Investigation (Free full text). 81 (6): 1710-1715. doi:10.1172/ ... "Genetic heterogeneity in the cerebrohepatorenal (Zellweger) syndrome and other inherited disorders with a generalized ...

*Solute carrier family 35 (CMP-sialic acid transporter), member A1

2005). "Genetic complementation reveals a novel human congenital disorder of glycosylation of type II, due to inactivation of ... "Sensitization and testing of guinea pigs with nickel sulfate". Dermatologica. 152 (6): 321-30. doi:10.1159/000251278. PMID ...

*Forward genetics

Once mutagenized and screened, typically a complementation test is done to ensure that mutant phenotypes arise from the same ... A genetic map can then be created using linkage and genetic markers, and then the gene of interest can be cloned and sequenced ... Cystic fibrosis however demonstrates how the process of forward genetics can elucidate a human genetic disorder. Genetic- ... Discovering disease loci using old forward genetic techniques was a very long and difficult process and much of the work went ...

*Evolution of sexual reproduction

The repair and complementation hypothesis assumes that genetic recombination is fundamentally a DNA repair process, and that ... Since hypotheses for the origins of sex are difficult to test experimentally (outside of Evolutionary computation), most ... These mutations are passed onto the next generation because the offspring are exact genetic clones of their parent. The genetic ... Organisms need to replicate their genetic material in an efficient and reliable manner. The necessity to repair genetic damage ...

*Paul Nurse

Lee, M. G.; Nurse, P. (1987). "Complementation used to clone a human homologue of the fission yeast cell cycle control gene ... Nurse, P.; Thuriaux, P.; Nasmyth, K. (1976). "Genetic control of the cell division cycle in the fission yeast ... for the constant testing of ideas." Furthermore, Nurse feels that scientific leaders "have a responsibility to expose the ...

*ERCC4

... and this gene restored UV resistance to cells of complementation group 4. Reflecting this cross-species genetic complementation ... The colon crypts are shaped like microscopic thick walled test tubes with a central hole down the length of the tube (the crypt ... Lee E, Levine EA, Franco VI, Allen GO, Gong F, Zhang Y, Hu JJ (2014). "Combined genetic and nutritional risk models of triple ... Busch D, Greiner C, Lewis K, Ford R, Adair G, Thompson L (Sep 1989). "Summary of complementation groups of UV-sensitive CHO ...

*BRCA1

Robert Cook-Deegan, MD et al (2010) Impact of Gene Patents and Licensing Practices on Access to Genetic Testing for Inherited ... Complementation Group S, a genetic disease associated with hypersensitivity to DNA crosslinking agents. BRCA1 is part of a ... Legal decisions surrounding the BRCA1 and BRCA2 patents will affect the field of genetic testing in general. A June 2013 ... Methods to test for the likelihood of a patient with mutations in BRCA1 and BRCA2 developing cancer were covered by patents ...

*APOBEC1

Deletion tests with mutant strands have shown that residues 181 to 210 are integral to mRNA editing, and there is most likely a ... CUGBP2 modulates C to U editing of apolipoprotein B mRNA by interacting with apobec-1 and ACF, the apobec-1 complementation ... "Complete phenotypic characterization of apobec-1 knockout mice with a wild-type genetic background and a human apolipoprotein B ... Tests involving A1 mutants with various deleted amino acid sequences have shown that editing activity is dependent on residues ...

*Heterosis

The genetic overdominance hypothesis states that some combinations of alleles (which can be obtained by crossing two inbred ... the steady rise in IQ test scores around the world during the twentieth century. However, James R. Flynn has pointed out that ... see Complementation (genetics)). It attributes the poor performance of inbred strains to the expression of homozygous ... When a population is small or inbred, it tends to lose genetic diversity. Inbreeding depression is the loss of fitness due to ...

*Epistasis

In this regression, the observed two locus genetic effects are treated as dependent variables and the "pure" genetic effects ... This is sometimes called allelic complementation, or interallelic complementation. It may be caused by several mechanisms, for ... In addition, in those tests which used artificial gene networks, negative epistasis is only found in more densely connected ... Quantitative genetics focuses on genetic variance due to genetic interactions. Any two locus interactions at a particular gene ...

*ENU

... test) stock that was used in genetic screens for testing mutagens such as radiations and chemicals. The T-stock mouse harbors 7 ... then this leads to non-allelic non-complementation. In a non-complementation screen, an ENU-induced male is crossed with a ... ENU is used as a genetic tool by designing a variety of genetic screens suitable to the researchers' interests. Depending on ... Genome-wide screens are most often useful for studying genetic diseases in which multiple genetic and biochemical pathways may ...

*Tunicate

... genetic complementation) and the avoidance of inbreeding depression. Botryllus schlosseri (class Ascidiacea) is a colonial ... The body of an ascidiacean is surrounded by a test or tunic, from which the subphylum derives its name. This varies in ... When, in 1845, Carl Schmidt first announced the presence in the test of some ascidians of a substance very similar to cellulose ...

*Heterozygote advantage

Because the genetic disorder is incompletely recessive, a person with only one SCA allele and one unaffected allele will have a ... The latter claim has been tested in an experiment, which showed outbreeding mice to exhibit MHC heterozygosity enhanced their ... found that the majority of cases of heterozygote advantage were due to complementation (or dominance), the masking of ... It is the most common genetic disease among people of European descent. The presence of a single CF mutation may influence ...

*Enterobacteria phage T4

Complementation, deletion, and recombination tests can be used to map out the rII gene locus by using T4. These bacteriophage ... HARM W (1958). "Multiplicity reactivation, marker rescue, and genetic recombination in phage T4 following x-ray inactivation". ... A Slot Machine, A Broken Test Tube: An Autobiography. Harper & Row, New York: 1984. Pp. 228. ISBN 0-06-015260-5 (USA and Canada ... LURIA SE, DULBECCO R (1948). "Lethal mutations, and inactivation of individual genetic determinants in bacteriophage". Genetics ...

*Bare lymphocyte syndrome type II

The diagnosis for Bare lymphocyte syndrome type II can be done via genetic testing A blood test could indicate decreased CD4+ T ... complementation group A - Conditions - GTR - NCBI". www.ncbi.nlm.nih.gov. Retrieved 2017-07-28. Matheux, Franck; Villard, Jean ... Bare lymphocyte syndrome type II (BLS II) is a rare recessive genetic condition in which a group of genes called major ... Review Ting, Jenny Pan-Yun; Trowsdale, John (April 2002). "Genetic Control of MHC Class II Expression". Cell. 109 (2): S21-S33 ...

*STAT1

When tested from whole blood, monocytes do not respond to BCG and IFNg doses with IL-12 production. In complete recessive form ... There are two main genetic impairments that can cause response to interferons type I and III. First there can be autosomal ... complementation group C, GNB2L1, IFNAR2, IRF1, ISGF3G Interleukin 27 receptor, alpha subunit, MCM5, Mammalian target of ... With various genomic and genetic methods was discovered, that a heterozygous gain of function mutation of STAT1 is a cause of ...

*Evolution by gene duplication

The IAD model have been previously tested in the lab by using an bacterial enzyme with dual function as starting point. This ... Hittinger CT, Carroll SB (2007). "Gene duplication and the adaptive evolution of a classic genetic switch". Nature. 449 (7163 ... "duplication-degeneration-complementation". This model was first introduced by Force et al. 1999. The first step is gene ... degeneration and complementation. Pseudogenes Molecular evolution Gene duplication Functional divergence Mutation Susumu Ohno ( ...

*Zebrafish

In another study, by White and Zon, an effort was made to therapeutically target the genetic program present in the tumor's ... Researchers frequently amputate the dorsal and ventral tail fins and analyze their regrowth to test for mutations. It has been ... it is not always easy to silence the activity one of the two gene paralogs reliably due to complementation by the other paralog ... Due to their short lifecycles and relatively large clutch sizes, zebrafish are a useful model for genetic studies. A common ...

*Lac operon

... in which genes or gene clusters are tested pairwise, is called a complementation test. This test is illustrated in the figure ( ... Gene regulation of the lac operon was the first genetic regulatory mechanism to be understood clearly, so it has become a ... In panel (e) the complementation test for repressor is shown. If one copy of the lac genes carries a mutation in lacI, but the ... and then designed their complementation tests to show this. The dominance of operator mutants also suggests a procedure to ...

*FANCA

It belongs to the Fanconi anaemia complementation group (FANC) family of genes of which 12 complementation groups are currently ... The primary diagnostic test for Fanconi anaemia is based on the increased chromosomal breakage seen in afflicted cells after ... Joenje H, Patel KJ (June 2001). "The emerging genetic and molecular basis of Fanconi anaemia". Nat. Rev. Genet. 2 (6): 446-57. ... Fanconi anaemia, complementation group A, also known as FAA, FACA and FANCA, is a protein which in humans is encoded by the ...
population and evolutionary genetics. My research uses evolutionary and population genetic theory as a framework for understanding the evolutionary significance of mutation rates and mutational phenomena.. Because the ultimate source of genetic variation is mutation, the evolution of mutation rates is a subject of basic interest in genetics. Considerable health implications exist as well: Recent findings have linked high somatic mutation rates with certain cancers, and high mutation rates have also been linked to pathogenicity in E. coli and Salmonella. Defective methyl-directed mismatch repair (hereafter, MMR) is implicated as the underlying mechanistic basis for high mutation rates in both of these cases. However, the basis for the evolutionary success of MMR-defective alleles remains to be examined rigorously. I am currently studying experimental populations of the bacterium Escherichia coli in which strikingly elevated general mutation rates have evolved. Genetic complementation analyses ...
dna dna polymerase radioisotope virus dna adenovirus dna replication dna sequence genetic engineering heredity nonhuman Adenoviruses Human Base Sequence Cell Nucleus DNA Viral DNA Directed DNA Polymerase Genes Viral Genetic Complementation Test Hela Cells Human Mutation Plasmids Virus ...
Blotting; Northern, Cell Division, Centrifugation; Density Gradient, Escherichia coli/metabolism, Genetic Complementation Test, Immunoblotting, Mutation, Protein Binding, Protein Biosynthesis, RNA; Bacterial/*chemistry, RNA; Messenger/metabolism, RNA-Binding Proteins/metabolism/*physiology, Research Support; Non-U.S. Govt, Ribosomes/*chemistry/metabolism, Subcellular Fractions, Sucrose/pharmacology, Time Factors ...
Thirty-three temperature-sensitive mutations defective in the start event of the cell division cycle of Saccharomyces cereuisiae were isolated and subjected to preliminary characterization. Complementation studies assigned these mutations to four complementation groups, one of which, cdc28, has been described previously. Genetic analysis revealed that these complementation groups define single nuclear genes, unlinked to one another. One of the three newly identified genes, cdc37, has been located in the yeast linkage map on chromosome IV, two meiotic map units distal to hom2.-Each mutation produces stage-specific arrest of cell division at start, the same point where mating pheromone interrupts division. After synchronization at start by incubation at the restrictive temperature, the mutants retain the capacity to enlarge and to conjugate.. ...
In bacteria, the highly conserved RsmA/CsrA family of RNA-binding proteins functions as global posttranscriptional regulators acting on mRNA translation and stability. Through phenotypic complementation of an rsmA mutant in Pseudomonas aeruginosa, we discovered a family member, termed RsmN. Elucidation of the RsmN crystal structure and that of the complex with a hairpin from the sRNA, RsmZ, reveals a uniquely inserted alpha helix, which redirects the polypeptide chain to form a distinctly different protein fold to the domain-swapped dimeric structure of RsmA homologs. The overall beta sheet structure required for RNA recognition is, however, preserved with compensatory sequence and structure differences, allowing the RsmN dimer to target binding motifs in both structured hairpin loops and flexible disordered RNAs. Phylogenetic analysis indicates that, although RsmN appears unique to P. aeruginosa, homologous proteins with the inserted alpha helix are more widespread and arose as a consequence of ...
The ability of various B10 congenic resistant strains to respond to the alloantigen H-2.2 was tested. High and low antibody-producing strains were distinguished by their anti-H-2.2 hemagglutinating respones. However, these strains do not differ in their ability to respond to these antigenic differences in the mixed lymphocyte culture. The humoral response to the H-2.2 alloantigen was shown to be controlled by two interacting genes localized within the H-2 complex. Thus, F1 hybrids prepared between parental low responder strains could yield high level immune responses. In addition, strains bearing recombinant H-2 haplotypes were used to map the two distinct genes controlling the immune response. The alleles at each locus were shown to be highly polymorphic as evidenced by the asymmetric complementation patterns observed. The restricted interactions of specific alleles was termed coupled complementation. The significance of the results in the terms of mechanisms of Ir gene control are discussed. ...
Occasionally, multiple mutations of a single wild type phenotype are observed. The appropriate genetic question to ask is whether any of the mutations are in a single gene, or whether each mutations represents one of the several genes necessary for a phenotype to be expressed. The simplest test to distinguish between the two possibilities is the complementation test. The test is simple to perform --- two mutants are crossed, and the F1 is analyzed. If th e F1 expresses the wild type phenotype, we conclude each mutation is in one of two possible genes necessary for the wild type phenotype. When it is shown that shown genetically that two (or more) genes control a phenotype, the genes are said to form a complementation group. Alternatively, if the F1 does not express the wild type phenotype, but rather a mutant phenotype, we conclude that both mutations occur in the same gene.. These two results can be explained by considering the importance of genes to phenotypic function. If two separate genes ...
It is not unusual to have series of mutations that confer similar phenotypes and also map to a identical or similar location on a chromosome. In such cases, the practicing geneticist performs a complementation test to determine if the mutations are allelic (that is, in the same gene) or non-allelic. If the mutations are allelic there should be no complementation whereas you could recover the wild type phenotype (though complementation) if the two mutations are on different genes. The specifics of strain construction vary depending on the experimental organism. However, the basic strategy in all cases is to construct a double heterozygote and then examine the phenotype of this organism. As mentioned above, a wild-type phenotype indicates that the two mutations complement one another and are therefore in different genes. Conversely, a mutant phenotype suggests the mutations are allelic to one another (that is, they fail to complement). We will construct double hets with the dumpy mutation of ...
In order to find out if a mutation under study in a forward genetics project is likely to be a newly discovered mutation or is, perhaps, in a previously characterized gene, we will perform a complementation analysis. If our mutation and gene has been previously characterized, this complementation analysis might tell us the name of our gene of interest. It is not unusual to have series of mutations that confer similar phenotypes and also map to a identical or similar location on a chromosome. Complementation testing can determine if two mutations are allelic (that is, in the same gene) or non-allelic (in different genes but both causing the same phenotype). This is done by crossing a mutant with a series of reference strains. In our case, we will use several different reference mutant strains. All have a Dpy phenotype, but in each strain the gene responsible for the dumpy defect has been located to a different known region of a chromosome ...
Creative Biolabs supplies Protein-fragment Complementation Assay (PCA) service to detect protein-protein interactions (PPIs) in vivo or in vitro.
The temperature-sensitive prp24-1 mutation defines a gene product required for the first step in pre-mRNA splicing. PRP24 is probably a component of the U6 snRNP particle. We have applied genetic reversion analysis to identify proteins that interact with PRP24. Spontaneous revertants of the temperature-sensitive (ts) prp24-1 phenotype were analyzed for those that are due to extragenic suppression. We then extended our analysis to screen for suppressors that confer a distinct conditional phenotype. We have identified a temperature-sensitive extragenic suppressor, which was shown by genetic complementation analysis to be allelic to prp21-1. This suppressor, prp21-2, accumulates pre-mRNA at the non-permissive temperature, a phenotype similar to that of prp21-1. prp21-2 completely suppresses the splicing defect and restores in vivo levels of the U6 snRNA in the prp24-1 strain. Genetic analysis of the suppressor showed that prp21-2 is not a bypass suppressor of prp24-1. The suppression of prp24-1 by ...
I have analyzed the time course of phage PR4 protein synthesis and have identified at least 34 proteins present in phage infected cells not detected in uninfected control cultures. In addition, I have isolated a more extensive set of conditional-lethal nonsense mutants of this virus. This collection of mutants permitted the identification of seven additional phage PR4 gene products, including the terminal genome protein and an accessory lytic factor. The present collection of phage PR4 mutants has been assigned to 19 distinct genetic groups on the basis of genetic complementation tests and sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis of the proteins produced in mutant infected UV irradiated cells ...
Hand pollinations were used for all genetic crosses and detailed pedigree information is available upon request.. Genetic complementation tests between the ems97406 mutation and other mutations known to affect Pl′ were conducted using +/ems97406 plants as pistillate parent. Staminate parent genotypes, number of crosses, and number of progeny plants with specific anther phenotypes are as follows: rmr1-1/rmr1-1, 4, 87 ACS 1-4, 2 ACS 5-6; rmr2-1/rmr2-1, 4, 88 ACS 1-4; mop1-1/mop1-1, 3, 56 ACS 1-4. Tests with the ems98225 mutation were conducted using ems98225 homozygotes as staminate parents. Pistillate parent genotypes, number of crosses, and number of progeny plants with specific anther phenotypes are as follows: rmr1-1/+, 3, 38 ACS 1-4; rmr2-1/+, 2, 25 ACS 1-4; mop1-2/+, 2, 31 ACS 1-4.. For both RNase protection analyses and in vitro transcription reactions, progeny sets were generated segregating 1:1 for +/rmr6-1 and rmr6-1/rmr6-1 siblings. Plants of these two genotypes were clearly ...
The use of genetic mutants has been invaluable in discovering components of molecular pathways. One of the most successful examples is the elucidation of intracellular mediators and signal transducers, which contribute to an IFN response. For example, the tyk kinase, which is associated with the receptor for type I IFN, was first discovered with the use of a gene complementation approach made possible by the availability of mutant cell lines defective in their responses to type I IFN (46). Similarly, a number of mutant cell lines defective in either type I or II IFN signaling have been instrumental in confirming the functional roles of Janus kinases and STAT molecules in the IFN pathway (47, 48).. Along the same vein, four mutant cell lines, G1 to G4, were genetically chosen on the basis of their selective loss of IFN-γ-induced MHC class II expression while retaining expression of other IFN-γ-induced genes (1). Analyses of these and other mutant lines clearly indicate that the IFN-γ induction ...
The worlds first wiki where authorship really matters. Due credit and reputation for authors. Imagine a global collaborative knowledge base for original thoughts.
The worlds first wiki where authorship really matters. Due credit and reputation for authors. Imagine a global collaborative knowledge base for original thoughts.
The features of tRNALys,3 that dictate why human immunodeficiency virus exclu-sively selects this tRNA as the primer for initiation of reverse transcription is unknown. The post-transcriptional modification at nucleotide 58 in the T??C stem-loop could play a role during plus-strand synthesis to stop reverse transcriptase from re-copying the tRNA primer. Nucleotides 53 and 54 within the T??C stem loop of the tRNA have been shown to be important to form the complex between tRNA and the HIV-1 viral genome during initiation of reverse transcription. To further delineate the features of the T??C stem loop in tRNALys,3 in reverse transcription, we used a complementation system in which E.coli tRNALys,3 is provided in trans to a mutated HIV-1 genome in which the PBS is comple-mentary to this tRNA to ascertain the effects that different mutants in the T??C stem loop of tRNALys,3 have on complementation. Alteration of nucleotide 58 from A to U (A58U), T54G and TG5453CC all resulted in tRNALys,3 that was ...
To address the importance of signals generated by the clontypic T cell receptor (TCR), we have developed a model system in which the critical adaptor protein, SLP-76, is conditionally deleted at various times during an immune response. Lack of SLP-76 abrogates measureable TCR-generated signals in T lymphocytes. This system allows us to generate a normal immune response and then alter the signaling capacity in a defined population of memory T cells. Interestingly, continuous SLP-76 expression is required for the long-term maintenance of CD4+ but not CD8+memory T cells. Studies using a genetic complementation approach are currently underway to determine if the PI3 kinase/AKT, Ras/MAPK and NFkB pathways are crucial for CD4+ memory T cell maintenance ...
L h moglobinurie paroxystique nocturne (HPN) ou syndrome de Marchiafava-Micheli est une pathologie rare. Elle est due une mutation clonale acquise affectant les cellules souches h matopo tiques. Les manifestations cliniques sont variables, avec une fr quence accrue d h molyse.. L HPN est une maladie orpheline. La population concern e est surtout l adulte jeune. La pr valence exacte de cette maladie n est pas connue. L HPN r sulte d une mutation du g ne PIG-A (Phosphatidyl Inositol Glycan complementation class A). Cette mutation aboutit la production de cellules souches d ficientes en prot ines GPI ancr es. Deux de ces prot ines, CD55 et CD59, prot gent normalement les globules rouges de l action lytique du compl ment. Leur absence se traduit par une lyse cellulaire avec lib ration du contenu intracellulaire. Trois ph notypes sont d crits : ph notype I = cellules normales ; ph notype II = d ficit partiel en prot ine GPI ; ph notype III = d ficit total. C est la proportion de cellules de ph notype ...
BioAssay record AID 1077914 submitted by ChEMBL: Antiviral activity against HIV1 infected in human Jurkat cells assessed as inhibition of viral replication by CAT gene-based transcomplementation assay.
Trans-complementation of ∆-NS1-WNV with ectopically expressed WNV NS1. A. Scheme for construction of ∆-NS1-WNV. Nucleotides 87 to 928 of the NS1 gene were d
The overall topic of this work is a graph operation known as edgelocal complementation (ELC) and its applications to iterative decoding of classical codes. Although these legacy codes are arguably not well-suited for graph-based decoding, they have other desirable properties resulting in much current research on the general problem of forging this alloy. From this perspective, these codes are typically referred to as highdensity parity-check codes. Our approach is to gain diversity by means of ELC. Based on the known link between ELC and the information sets of a code, C, we identify a one-to-one relationship between ELC operations and the automorphism group of a code, Aut(C). With respect to a specific parity-check matrix, H, we classify these code-preserving permutations into trivial and nontrivial permutations, based on whether the matrix is preserved (under ELC) up to row permutations, or not. The corresponding iso-ELC operations preserve the structure of the graph, and simulation data are ...
Complementation, as opposed to psychological integration, underlies a spiritual technique that requires a toning down of dominant consciousness.
Molecular characterization and protein analysis of the cap region, which is essential for encapsulation in Bacillus anthracis.: By using genetic complementation
Compounds are evaluated for their binding to naturally occurring receptors, by employing the natural ligand conjugated to an enzyme donor fragment of β-galactosidase for competing with the sample compound for the natural acceptor binding site or in the absence of competition where the sample compound binds to an allosteric site. By adding the enzyme acceptor fragment of the β-galactosidase and substrate, the binding affinity of the sample compound may be evaluated as a measure of agonist or antagonist capability.
The I-2 locus in tomato confers resistance to race 2 of the soil-borne fungus Fusarium oxysporum f sp lycopersici. The selective restriction fragment amplification (AFLP) positional cloning strategy was used to identify I-2 in the tomato genome. A yeast artificial chromosome (YAC) clone covering ~750 kb encompassing the I-2 locus was isolated, and the AFLP technique was used to derive tightly linked AFLP markers from this YAC clone. Genetic complementation analysis in transgenic R1 plants using a set of overlapping cosmids covering the I-2 locus revealed three cosmids giving full resistance to F. o. lycopersici race 2. These cosmids shared a 7-kb DNA fragment containing an open reading frame encoding a protein with similarity to the nucleotide binding site leucine-rich repeat family of resistance genes. At the I-2 locus, we identified six additional homologs that included the recently identified I-2C-1 and I-2C-2 genes. However, cosmids containing the I-2C-1 or I-2C-2 gene could not confer ...
In this study we demonstrated successful transcomplementation of KUN genomic RNAs with large in-frame deletions in the NS1 and NS3 genes by providing corresponding helper proteins from KUN replicon RNA persistently replicating in repBHK cells. Previously we showed trans complementation of KUN genomic RNAs with C-terminal deletions of more than half of the NS5 gene (23). By combining these individual deletions in the same RNA molecule, we were able to demonstrate trans complementation of RNAs containing double deletions in the NS1 and NS5 genes or triple deletions in the NS1, NS3, and NS5 genes. This is the first demonstration of trans complementation of replication of flavivirus RNAs containing deletions of as much as 84 to 97% of the NS1 gene, or of any deletion in the NS3 gene, or of deletions in two or three NS genes in the same RNA molecule.. In this and our previous studies we have attempted complementation of deletions introduced into over 80% of the nonstructural region of the infectious ...
MHC‐II deficiency is a genetic disease of gene regulation. It is due to defects in regulatory factors that are essential for both constitutive and IFN‐γ inducible expression of MHC‐II genes (Reith et al., 1995, 1997; Mach et al., 1996). Together with a number of in vitro generated regulatory mutants, MHC‐II deficiency patients have been classified into at least four different complementation groups (A, B, C and D) believed to correspond to at least four distinct regulatory genes (Hume and Lee, 1989; Benichou and Strominger, 1991; Seidl et al., 1992; Lisowska‐Grospierre et al., 1994). The disease thus provides a genetic approach to identify genes encoding several of the trans‐acting regulatory factors involved and therefore represents an ideal model system for the dissection of the molecular mechanisms controlling transcriptional activation of MHC‐II genes. The relevant regulatory genes can be identified on the basis of a powerful functional criterion, namely the ability to ...
Cell division in Chlamydiae is poorly understood as apparent homologs to most conserved bacterial cell division proteins are lacking and presence of elongation (rod shape) associated proteins indicate non-canonical mechanisms may be employed. The rod-shape determining protein MreB has been proposed as playing a unique role in chlamydial cell division. In other organisms, MreB is part of an elongation complex that requires RodZ for proper function. A recent study reported that the protein encoded by ORF CT009 interacts with MreB despite low sequence similarity to RodZ. The studies in this paper expand on those observations through protein structure, mutagenesis and cellular localization analyses. Structural analysis indicated that CT009 shares high level of structural similarity to RodZ, revealing the conserved orientation of two residues critical for MreB interaction. Substitutions eliminated MreB protein interaction and partial complementation provided by CT009 in RodZ deficient Escherichia ...
Cell fusion is known to underlie key developmental processes in humans and is postulated to contribute to tissue maintenance and even carcinogenesis. The mechanistic details of cell fusion, especially between different cell types, have been difficult
For instance they could do some screens for temperature-sensitive mutants (huge, massive saunas in action). Imagine the figures in the papers to go along with this sort of experiments. Some allele crossing experiments in search of synthetic lethality - that would be great as well. With photos of F0 and F1. Auxotrophic humans with plasmids complementing their deficiency as useful tools - complementation experiments will be particularly cruel - no complementation - well, tough luck ...
Cellular biochemical machineries, what we call pathways, consist of dynamically assembling and disassembling macromolecular complexes. Although our models for the organization of biochemical machines...
Molecular Cloning, also known as Maniatis, has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential.
Bacteria have evolved a wide range of chemoreceptors with different ligand specificities. Typically, chemoreceptors bind ligands with elevated specificity and ligands serve as growth substrates. However, there is a chemoreceptor family that has a broad ligand specificity including many compounds that are not of metabolic value. To advance the understanding of this family, we have used the PcaY_PP (PP2643) chemoreceptor of Pseudomonas putida KT2440 as a model. Using Isothermal Titration Calorimetry we showed here that the recombinant ligand binding domain (LBD) of PcaY_PP recognizes 17 different C6-ring containing carboxylic acids with KD values between 3.7 and 138 µM and chemoeffector affinity correlated with the magnitude of the chemotactic response. Mutation of the pcaY_PP gene abolished chemotaxis to these compounds; phenotype that was restored following gene complementation. Growth experiments using PcaY_PP ligands as sole C-sources revealed functional relationships between their metabolic
Poliovirus RNA replicates in membrane-associated replication complexes in the cytoplasm of infected cells. By using a reversible inhibitor of poliovirus RNA replication, it is possible to synchronize viral RNA replication. The processing of the viral polyprotein results in the formation of the individual viral proteins along with stable intermediates in the processing pathway. To expand the utility of the in vitro complementation assay, experiments were designed to determine if all of the viral replication proteins could be provided in trans to support the replication of mutant RNA templates. The authors engineered two transcript RNAs (DJB2 and DJB15) that contained large out-of-frame deletions in the polyprotein coding sequence. The results to date using the in vitro complementation assay indicate that the 5 cloverleaf, the 3 nontranslated region (NTR), and the poly(A) tail are the minimum sequences required for negative-strand synthesis. Previous studies have shown that the 5 cloverleaf plays an
The dynamic assembly of multiprotein complexes is a central mechanism of many cell signaling pathways. This process is key to maintaining the spatiotemporal specificity required for an accurate, yet adaptive, response to rapidly changing cellular conditions. We describe a technique for the specific isolation and downstream proteomic characterization of any two interacting proteins, to the exclusion of their individual moieties and competing binding partners. We termed the approach bimolecular complementation affinity purification (BiCAP) because it combines the use of conformation-specific nanobodies with a protein-fragment complementation assay with affinity purification. Using BiCAP, we characterized the specific interactome of the epidermal growth factor receptor (EGFR) family member ERBB2 when in the form of a homodimer or when in the form of a heterodimer with either EGFR or ERBB3. We identified dimer-specific interaction patterns for key adaptor proteins and identified a number of previously
Author Summary Many viruses encode homologs of human oncogenes, including the gammaherpesvirus viral cyclin genes. These viruses cause lifelong infection associated with chronic diseases, including malignancies, which are exacerbated in immune deficiency. The conserved viral cyclins were first recognized nearly two decades ago, and despite extensive interest and study, their essential features for virus infection and disease have been elusive. We used a mouse model of these viruses to make recombinant viruses with viral or human cyclins knocked into the endogenous locus. We then determined the requirements for cyclins by genetic complementation in three distinct viral cyclin dependent aspects of infection. We report that the viral cyclins of different gammaherpesviruses are able to support all three stages of infection. However, none of the human cyclins can, and instead comprise distinct complementation groups that are functional in non-overlapping aspects of infection. We showed that gammaherpesvirus
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This report describes mutants of the zebrafish having phenotypes causing a general arrest in early morphogenesis. These mutants identify a group of loci making up about 20% of the loci identified by mutants with visible morphological phenotypes within the first day of development. There are 12 Class I mutants, which fall into 5 complementation groups and have cells that lyse before morphological defects are observed. Mutants at three loci, speed bump, ogre and zombie, display abnormal nuclei. The 8 Class II mutants, which fall into 6 complementation groups, arrest development before cell lysis is observed. These mutants seemingly stop development in the late segmentation stages, and maintain a body shape similar to a 20 hour embryo. Mutations in speed bump, ogre, zombie, specter, poltergeist and troll were tested for cell lethality by transplanting mutant cells into wild-type hosts. With poltergeist, transplanted mutant cells all survive. The remainder of the mutants tested were autonomously but ...
... 2 family (SnRK2) are crucial in mediating different stress-adaptive responses. threonine due to systematic adjustments in the flanking amino acidity sequence. Our outcomes designate the ABA-responsive-element Binding Element 3 (ABF3), which settings area of the ABA-regulated transcriptome, as an authentic OST1 substrate. Bimolecular Fluorescence Complementation experiments indicate that ABF3 interacts with OST1 in the nuclei of living plant cells directly. which phospho-T451 is very important to stabilization of ABF3. Conclusions/Significance Altogether, our results claim that OST1 phosphorylates ABF3 on T451 to make a 14-3-3 binding theme. Inside a wider physiological framework, we suggest that the future reactions to 507475-17-4 manufacture ABA that want sustained gene manifestation is, partly, mediated from the stabilization of ABFs powered by ABA-activated SnRK2s. Intro The vegetable hormone abscisic ...
1DP0: High resolution refinement of beta-galactosidase in a new crystal form reveals multiple metal-binding sites and provides a structural basis for alpha-complementation.
1DP0: High resolution refinement of beta-galactosidase in a new crystal form reveals multiple metal-binding sites and provides a structural basis for alpha-complementation.
MORAN, JAMES PAUL, "POLAR EFFECTS ON THE RATES OF FORMATION AND DIMERIZATION OF FREE RADICALSFROM ETHYL ACETATE" (1963). Doctoral Dissertations. AAI6403549 ...
XPA antibody (xeroderma pigmentosum, complementation group A) for ICC/IF, IHC-P, IP, WB. Anti-XPA pAb (GTX103168) is tested in Human samples. 100% Ab-Assurance.
XPA antibody [12F5] (xeroderma pigmentosum, complementation group A) for ICC/IF, IHC-P, WB. Anti-XPA mAb (GTX72316) is tested in Human samples. 100% Ab-Assurance.
All steps were performed in a darkroom under dim red light. RPE/choroid samples were removed from eyecups and homogenized in 200 μL PBS (pH 7.4, 150 mM NaCl, 1.06 mM KH2PO4, 5.60 mM Na2HPO4) in a disposable homogenizer. Homogenate (10 μL) was taken for the protein assay (Microplate Pierce Coomassie [Bradford] Plus Protein Assay; Thermo Fisher Scientific Inc., Rockford, IL). The remaining homogenate was transferred to a 15 mL screw-top polypropylene tube (BD Falcon; BD Biosciences, San Jose, CA), and 400 μL methanol was added. Retinoids were extracted two times by adding 1.5 mL hexane and vortex mixing at top speed for 1 minute. Samples were centrifuged at 1424g for 3 minutes. Hexane was drawn off the upper phase and transferred to a fresh 15 mL screw-top polypropylene tube (BD Falcon; BD Biosciences). Pooled hexane extracts were dried in an evaporation system (TurboVap LV; Zymark Corporation, Hopkinton, MA). Samples were redissolved in 100 μL hexane by vortex mixing for 1 minute followed by ...
Protein fragment complementation assays for high-throughput and high-content screening - The present invention provides protein fragment complementation assays for drug discovery, in particular to identify compounds that activate or inhibit cellular pathways. Based on the selection of an interacting protein pair combined with an appropriate PCA reporter, the assays may be run in high-throughput or high-content mode and may be used in automated screening of libraries of compounds. The interacting pair may be selected by cDNA library screening; by gene-by-gene interaction mapping; or by prior knowledge of a pathway. Fluorescent and luminescent assays can be constructed using the methods provided herein. The selection of suitable PCA reporters for high-throughput or high-content (high-context) assay formats is described for a diversity of reporters, with particular detail provided for examples of monomeric enzymes and fluorescent proteins. Methods are described for constructing such assays for one ...
... (Gr. ?????? "to increase"; ????? "nourishment") is the inability of an organism to synthesize a particular organic compound required for its growth (as defined by IUPAC). An auxotroph is an organism that displays this characteristic; auxotrophic is the corresponding adjective. Auxotrophy is the opposite of prototrophy, which is characterized by the ability to synthesize all the compounds needed for growth. The method of replica plating implemented by Esther Lederberg included auxotrophs that were temperature-sensitive; that is, their ability to synthesize was temperature-dependent. (Auxotrophs are usually not temperature-dependent. They can also depend on other factors, such as light intensity or wavelength.) Multiple auxotrophs can also coexist at the same time, within the same organism.[1] In genetics, a strain is said to be auxotrophic if it carries a mutation that renders it unable to synthesize an essential compound. For example, a yeast mutant with an inactivated uracil ...
COMMENT We thank our colleagues for taking the time to read our paper and comment. Dr. Almeidas points are well taken; it is possible that there is some interference of normal trafficking after complementation (as we pointed out in the manuscript and in our .... ...
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A genetic analysis of thiamine metabolism has been carried out in the budding yeast, Saccharomyces cerevisiae. A collection of thiamine auxotrophic mutants were isolated following UV and Ty insertion mutagenesis. The mutations responsible for the auxotrophic phenotypes were characterised to different extents through complementation analysis, molecular cloning and enzyme assays. In total 171 mutants were analysed and all of these have been assigned to complementation groups, genes and/or functions. Some newly isolated mutations were found to be allelic with the known biosynthetic genes, THI4 and THI6 others were in the regulatory genes, THI2 and THI3 two more defined a new function for the transcription factor, Pdc2p, namely thiamine gene activation. In addition the previously known mutations, thil, thi2, and thi3, were complemented and the sequences of the wild-type THI1, THI2 and THI3 genes were found. From the deduced amino acid sequences roles for the gene products were hypothesised. The ...
The completion of the entire sequence of Mycobacterium tuberculosis (Cole et al., 1998) launched a new era in tuberculosis research. In order to study the function of M. tuberculosis genes, several mutants have been produced by homologous recombination and studied in animal models (Parish and Stoker 2000; Movahedzadeh et al., 2004, 2008, 2010). There is a wide range of phenotypes, from highly attenuated mutants (Smith et al., 2001; Movahedzadeh et al., 2004) to hypervirulent strains (McAdam et al., 2002; Parish et al., 2003). These phenotypes require confirmation by the generation of complementation strains, whereby the wild-type copy of the gene is re-introduced into the mutant strain. By complementation of the mutant strain, one can ensure that the observed mutant phenotype, e.g. increased virulence of M. tuberculosis with the loss of dosR, is actually due to the loss of dosR and not to secondary mutations that may have arisen during the creation of the mutant strain.. Several cloning systems ...
Thirty-seven nonhemolytic/nonbacteriocinogenic mutations in Enterococcus (Streptococcus) faecalis plasmid pAD1 were generated by Tn917 insertion. All were found to belong to one of two complementation classes. Each class of mutants secreted either hemolysin/bacteriocin (Hly/Bac) component A or L into the culture medium. DNA encoding Hly/Bac was cloned in Escherichia coli in which both components of the hemolysin were expressed individually and collectively. The region encoding components A and L was further defined by deletion analysis and physically mapped. A total of approximately 8.4 kilobases of pAD1 DNA were observed to be required for hemolysin expression. Hly/Bac activity of the wild-type and the inactive L substance was observed to be heat stable. Active Hly/Bac resulting from incubating separately secreted components A and L was also found to be heat stable. The results indicate that component A activates component L and that activated component L possesses the Hly/Bac activity. ...
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Reaktivität: Hund, Pferd, Human and more. 116 verschiedene ERCC4 Antikörper vergleichen. Alle direkt auf antikörper-online bestellbar!
Historians believe that in newspost ,20020424171001.01655.00002368 at mb-cl.aol.com, on Wed, 24 Apr 2002, JSUPolek ,jsupolek at aol.com, penned the following literary masterpiece: ,Does anyone know of a high copy number vector that does not contain the ,truncated lacZ gene? Ive been using pBR322 pHC624, pHC312, pHC314. Single point mutation of a pBR322 derivative (its a tet minus derivative as tet is lethal at high copy). Published many many years ago in Gene by a Hungarian group I think. Duncan -- I love deadlines. I especially like the whooshing noise they make as they go flying by. Duncan Clark GeneSys Ltd ...
Homo sapiens excision repair cross-complementing rodent repair deficiency, complementation group 8 (ERCC8), transcript variant 2, mRNA. (H00001161-R02) - Products - Abnova
Using a selection system that was enriched for mutants unable to grow on low-iron media, Askwith et al. reported the identification of a mutant, fet3, that was unable to grow on low-iron media (14). This mutant had a normal surface reductase activity but was unable to accumulate 59Fe. A gene that could complement both the low-iron growth defect and the inability to accumulate radioactive iron was identified by complementation of the mutant strain with a genomic library. Genetic studies.... ...
Li Zhang is the author of these articles in the Journal of Visualized Experiments: Højeffektiv generation af antigen-specifikke primære mus cytotoksiske T-celler til funktionel testning i en autoimmun diabetes model, Måling af hæmoglobinsyntese Niveauer i pattedyrceller, Farvning protokoller for Menneskerettigheder bugspytkirteløer, Detection of Protein Interactions in Plant using a Gateway Compatible Bimolecular Fluorescence Complementation (BiFC) System
Complete information for TS13 gene (Genetic Locus), Temperature Sensitivity Complementation, Ts13, including: function, proteins, disorders, pathways, orthologs, and expression. GeneCards - The Human Gene Compendium
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Cloning of Clostridium thermocellum acetate kinase (ack) and/or phosphotransacetylase (pta) genes in Escherichia coli by functional complementation of ack and/or pta mutants was complicated by an alternative acetate assimilation pathway involving acetyl-CoA synthetase (ACS). In addition to the problems encountered with the complementation approach, cloning of these genes was not readily achieved using heterologous probing with corresponding genes from Escherichia coli and Methanosarcina thermophila due to the lack of sufficient homology. The use of a PCR-based approach, on the other hand, yielded a specific C. Thermocellum gene fragment which showed significant sequence identity to the ack gene for which primers were designed. The subcloned ack fragment was then successfully used as a probe for the isolation of the corresponding gene and restriction analysis of that region.
CDC23 is required in Saccharomyces cerevisiae for cell cycle progression through the G2/M transition. The CDC23 gene product contains tandem, imperfect repeats, termed tetratricopeptide repeats, (TPR) units common to a protein family that includes several other nuclear division CDC genes. In this report we have used mutagenesis to probe the functional significance of the TPR units within CDC23. Analysis of truncated derivatives indicates that the TPR block of CDC23 is necessary for the function or stability of the polypeptide. In-frame deletion of a single TPR unit within the repeat block proved sufficient to inactivate CDC23 in vivo, though this allele could rescue the temperature-sensitive defect of a cdc23 point mutant by intragenic complementation. By both in vitro and in vivo mutagenesis techniques, 17 thermolabile cdc23 alleles were produced and examined. Fourteen alleles contained single amino acid changes that were found to cluster within two distinct mutable domains, one of which ...
The AINTEGUMENTA-LIKE6/PLETHORA3 (AIL6/PLT3) gene of Arabidopsis thaliana is a key regulator of growth and patterning in both shoots and roots. AIL6 encodes an AINTEGUMENTA-LIKE/PLETHORA (AIL/PLT) transcription factor that is expressed in the root stem cell niche, the peripheral region of the shoot apical meristem and young lateral organ primordia. In flowers, AIL6 acts redundantly with AINTEGUMENTA (ANT) to regulate floral organ positioning, growth, identity and patterning. Experiments were undertaken to define the genomic regions required for AIL6 function and expression in flowers. Transgenic plants expressing a copy of the coding region of AIL6 in the context of 7.7 kb of 5′ sequence and 919 bp of 3′ sequence (AIL6:cAIL6-3′) fail to fully complement AIL6 function when assayed in the ant-4 ail6-2 double mutant background. In contrast, a genomic copy of AIL6 with the same amount of 5′ and 3′ sequence (AIL6:gAIL6-3′) can fully complement ant-4 ail6-2. In addition, a genomic copy of AIL6
The stabilisation of acetylcholine receptors (AChRs) at the neuromuscular junction depends on muscle activity and the cooperative action of myosin Va and protein kinase A (PKA) type I. To execute its function, PKA has to be present in a subsynaptic microdomain where it is enriched by anchoring proteins. Here, we show that the AChR-associated protein, rapsyn, interacts with PKA type I in C2C12 and T-REx293 cells as well as in live mouse muscle beneath the neuromuscular junction. Molecular modelling, immunoprecipitation and bimolecular fluorescence complementation approaches identify an α-helical stretch of rapsyn to be crucial for binding to the dimerisation and docking domain of PKA type I. When expressed in live mouse muscle, a peptide encompassing the rapsyn α-helical sequence efficiently delocalises PKA type I from the neuromuscular junction. The same peptide, as well as a rapsyn construct lacking the α-helical domain, induces severe alteration of acetylcholine receptor turnover as well as
Fungal strains deficient in the non-homologous end-joining (NHEJ) pathway are excellent recipient strains for gene targeting approaches. In addition, NHEJ-deficiency can facilitate the formation of heterokaryons which allows rapid identification of essential genes. However, the use of NHEJ-deficient strains can also pose some limitations for gene function analyses. For example, lack of the NHEJ pathway can interfere with phenotypic analyses and complicate complementation studies. Moreover, heterokaryons are difficult to propagate and re-transform. We describe here strategies and methods to circumvent these problems and to better exploit the power of NHEJ-deficient strains. We provide methods for the establishment of transiently deficient NHEJ strains, for improved complementation analyses using AMA1-based vectors and for fast identification and propagation of heterokaryons. The methods described are applicable for a wide range of filamentous fungi ...
The putative role of the S. cerevisiae vacuole in osmohomeostasis, as well as its biogenesis was analysed by taking a mutational approach. 97 mutants unable to tolerate high concentrations of salt were isolated and examined for aberrant vacuolar phenotypes. A comprehensive phenotypic analysis was able to demonstrate that apart from osmosensitivity most mutations conferred other properties such as altered vacuolar morphology, the inability to perform gluconeogenesis and/or the mislocalization of vacuolar proteins to the cell surface. The mutants fall into at least 20 complementation groups, termed ssv for salt sensitive vacuolar mutants, of which 3 genetically overlap with complementation groups isolated by others. This analysis provides evidence that in 5. cerevisiae correct vacuolar biogenesis is required for osmotolerance as well as other important cellular processes. To elucidate vacuolar osmohomeostasis at the molecular level, one gene, SSV7, was cloned from a genomic DNA library by ...
Tests for allelism among mice with four different mutant alleles at the shaker-with-syndactylism locus on mouse Chromosome (Chr) 18 provide evidence that the original radiation-induced mutation, sy, is a deletion including at least two genes associated with distinct phenotypes. Mice homozygous for sy have syndactylous feet and other skeletal malformations, are deaf, and exhibit abnormal behavior characteristic of vestibular dysfunction. Two less severe spontaneous mutations, shown to be allelic with sy, cause syndactylism when homozygous (hence named fused phalanges, sy(fp) and sy(fp-2J)), but do not affect hearing and behavior. Here we describe a third spontaneous mutation allelic with sy that does not affect foot morphology (hence named no syndactylism, sy(ns)), but that does cause deafness and balance defects when homozygous. Complementation test results indicate that sy(fp) and sy(fp-2J) are alleles of the same gene, but that sy(ns) is an allele of a different gene. The original sy
Dear yeast people, An INRA grant will be available in early 1995 for a postdoctoral position to work in the field of the regulation of the levels of cytokinins (plant hormones) in plants. The project involves the cloning of genes involved in the interconversion between cytokinins/purines bases, ribosides and nucleotides and in their catabolism by functional complementation of appropriate mutants of E. coli and/or yeasts. Candidates with training in microbiology, especially yeast genetics (why not in the area of purine utilization ?), and having an interest (or a potential interest) in the molecuar analysis of plant development are encouraged to postulate. Interested candidates should contact me first by Email for details. Dr Michel LALOUE INRA Laboratoire de Biologie Cellulaire Route de Saint-Cyr 78026 Versailles Cedex Fax : (33) 1 30 83 30 99 ...
ERCC5 - ERCC5 (untagged)-Human excision repair cross-complementing rodent repair deficiency, complementation group 5 (ERCC5) available for purchase from OriGene - Your Gene Company.
Sigma-Aldrich offers abstracts and full-text articles by [Duancheng Wen, Nestor Saiz, Zev Rosenwaks, Anna-Katerina Hadjantonakis, Shahin Rafii].
Dblog is the interwebositry for all things creative, inspirational and elegant from Ds world of art, design, cinema, T.V., comics, toys and pop culture. Drop me line or an interesting link any time ...
We have isolated three independent Chinese hamster ovary cell mutants (B3853, I223, and M311) with temperature-sensitive, pleiotropic defects in receptor-mediated endocytosis. Activities affected at 41 degrees C include uptake via the D-mannose 6-phosphate receptor, accumulation of Fe from diferric transferrin, uptake of alpha 2-macroglobulin, compartmentalization of newly synthesized acid hydrolases, resistance to ricin, and sensitivity to diphtheria and Pseudomonas toxins and modeccin. The three mutants also displayed decreased sialylation of some secreted glycoproteins at 41 degrees C, reminiscent of the nonconditional mutant DTG1-5-4 that showed both endocytic and Golgi-associated defects (Robbins, A.R., C. Oliver, J.L. Bateman, S.S. Krag, C.J. Galloway, and I. Mellman, 1984, J. Cell Biol., 99:1296-1308). Phenotypic changes were detectable within 30 min after transfer of the mutants to 41 degrees C; maximal alteration of most susceptible functions was obtained 4 h after temperature shift. At ...
0162]Atkins Latham, K. and R. S. Lloyd. T4 Endonuclease V. Perspectives on Catalysis. In DNA Damage-Effects on DNA Structure and Protein Recognition Annals of New York Academy of Science, volume 726, 1994. pp181-197. [0163]Augustine, M. L., R. W. Hamilton, M. L. Dodson and R. S. Lloyd. Oligonucleotide Site Directed Mutagenesis of All Histidine Residues within the T4 Endonuclease V Gene: Role in Enzyme-Nontarget DNA Binding. Biochemistry 30:8052-8059, 1991. [0164]Chenevert, J., L. Naumovski, R. Schultz, E. Friedberg. Partial Complementation of the UV Sensitivity of E. coli and Yeast Excision Repair Recombinants by the cloned deny gene of Bacteriophage T4. Molecular and General Genetics 203:163-171, 1986. [0165]Dodson, M. and R. Lloyd. Structure-Function Studies of the T4 endonuclease V Repair Enzyme. Mutation Research 218:49-65, 1989. [0166]Doi, T., A. Recktenwald, Y. Karaki, M. Kikuchi, K. Morikawa, M. Ikehara, T. Inaoka, N. Hori, E. Ohtsuka. Role of the Basic Amino Acid Cluster and Glu-23 in ...
HtrA is known to be an important stress response protease for many bacteria and has been shown to be critical for virulence in many bacteria, including intracellular pathogens Salmonella enterica and Legionella pneumophila [29, 30]. There is considerable evidence from both microarray and proteomic studies that HtrA is expressed in Chlamydia.. In the absence of a genetic manipulation system, a complementation approach was used to test the physiological function of C. trachomatis HtrA in a heterologous host (E. coli). E. coli HtrA protein (EcHtrA) and C. trachomatis HtrA protein (CtHtrA) are known to have differences in substrate specificity for their protease activities, although both have temperature activated protease activity, and are specific for unfolded proteins [4, 8]. The findings reported here show that the C. trachomatis htrA was able to protect E. coli htrA- against its lethal high temperature phenotype. This suggests that the ability to chaperone and degrade unfolded proteins, ...
Citation: N/A Interpretive Summary: Gibberella zeae causes wheat ear blight and also contaminates grain with the toxin deoxynivalenol. In this study, we conducted genetic analysis of the fungus and tested its virulence on wheat ears in the greenhouse. Our results show that the ability to produce deoxynivalenol increases the ability of this fungus to cause wheat ear blight. This information should help wheat breeders develop cultivars that are resistant to ear blight. Technical Abstract: Gibberella zeae causes wheat ear blight and produces trichothecene toxins in infected grain. In previous studies, trichothecene production in this fungus was disabled by specific disruption of the trichodiene synthase gene (Tri5) and was restored by two methods: gene reversion and transformation-mediated mutant complementation. In previous field tests of wheat ear blight, trichothecene-nonproducing mutants were less virulent than the wild-type progenitor strain from which they were derived. ...
A total of 62 out of 89 essential genes were successfully identified and 60 out of 62 were matched to their molecular counterpart. This large number demonstrates the value of our method. However, we were not able to identify any appropriate mutations in the sequences of the other 27 genes. Complementation tests and PCR sequencing were conducted to validate possible candidates for seven of those 27; however, we found that they were not essential genes. We do not know the reason(s) for this, but we hypothesize several possible reasons. First is strain mix-ups; however, this is unlikely because during the construction of the strains containing let-500 the terminal phenotypes of all the lethals were the same as noted when the lethals were first analyzed. Second is sequencing errors, as it is possible that there were not enough sequencing reads to support some of the lethal mutations. For example, only one out of 32 sequencing reads (3%) supports C to T change in unc-46(e177) in the let-417(s204) ...
RNA Detection in Live Bacterial Cells Using Fluorescent Protein Complementation Triggered by Interaction of Two RNA Aptamers with Two RNA-Binding Peptides. . Download books free in pdf. Online library with books, university works and thousands of documents available to read online and download.
Bacteria possess a number of cell-associated and secreted molecules, termed bacterial modulins, that stimulate the release of proinflammatory mediators in the host (4). In previous work, we (1, 3) and others (14) have demonstrated that isolated flagella or fragments of isolated flagella from gram-negative bacteria elicit the production of TNF-α in cultures of adherent human PBMC and monocyte-like cell lines. Genetic complementation in afliC deletion mutant identified flagellin as the key component of the flagella that was essential for the induction of cytokine synthesis (1). Although flagella from other gram-negative organisms, such as E. coli, P. aeruginosa, and Y. enterocolitica, also stimulated TNF-α synthesis by human monocytes, flagella fromSalmonella strains were generally the most potent inducers (1).. In the present study, we demonstrate that purifiedSalmonella FliC and FljB are exceptionally potent inducers of TNF-α synthesis, with detectable amounts of TNF-α being induced in cells ...
ERCC2 - ERCC2 Mutant (L485P), Myc-DDK-tagged ORF clone of Homo sapiens excision repair cross-complementing rodent repair deficiency, complementation group 2 (ERCC2), transcript variant 1 as transfection-ready DNA available for purchase from OriGene - Your Gene Company.
Reversible janus associated kinase (JAK) inhibitors such as tofacitinib and decernotinib block cytokine signaling and are efficacious in treating autoimmune diseases. However, therapeutic doses are limited due to inhibition of other JAK/signal transducer and activator of transcription pathways associated with hematopoiesis, lipid biogenesis, infection, and immune responses. A selective JAK3 inhibitor may have a better therapeutic index; however, until recently, no compounds have been described that maintain JAK3 selectivity in cells, as well as against the kinome, with good physicochemical properties to test the JAK3 hypothesis in vivo. To quantify the biochemical basis for JAK isozyme selectivity, we determined that the apparent Km value for each JAK isozyme ranged from 31.8 to 2.9 μM for JAK1 and JAK3, respectively. To confirm compound activity in cells, we developed a novel enzyme complementation assay that read activity of single JAK isozymes in a cellular context. Reversible JAK3 ...
This line is a derivative of the CHO-K1 cell line (see ATCC CCL-61). UV24 is a UV sensitive line derived from AA8 (see ATCC CRL-1859).
Get an answer for Name those parts of a flower which serve the same function as the following do in the animal:- 1. Testis 2. Ovary 3. Eggs 4. Sperms and find homework help for other Science questions at eNotes
We provided in vivo and in vitro data indicating that D5 is a DNA primase. First, a D5 ts mutant complementation assay (13) demonstrated the importance of conserved amino acids in the predicted primase active site for DNA replication. Furthermore, purified recombinant D5 exhibited primase activity that depended on conserved amino acids in the predicted active site.. Primases are DNA-dependent RNA polymerases that synthesize oligoribonucleotides 2-15 nt or longer, usually starting with ATP or GTP (18). Generally, any single-stranded DNA can serve as a template, although there may be preferential usage of some sequences. D5 primase activity was demonstrated by using single-stranded circular φX174 and M13 phage templates. A discrete RNase-sensitive band migrated near the 14-nt marker. We cannot be sure of the actual length of this oligoribonucleotide, because the markers were phosphorylated, and small oligonucleotides migrate anomalously in high percentage polyacrylamide gels (33). However, the ...
Complete information for FANCB gene (Protein Coding), Fanconi Anemia Complementation Group B, including: function, proteins, disorders, pathways, orthologs, and expression. GeneCards - The Human Gene Compendium
John Robert Stanley Fincham FRS FRSE (11 August 1926 - 9 February 2005) was a noted British geneticist who made important contributions to biochemical genetics and microbial genetics. Fincham was educated at Peterhouse, Cambridge, where he read Natural Sciences. He earned his PhD in the Botany School at Cambridge and then did a years postgraduate research at the California Institute of Technology with Sterling Emerson (whose daughter Ann he married). Fincham laboratory was among the first to demonstrate "intragenic complementation" through finding "pseudowild" progeny from am1 × am2 crosses. He obtained the first direct evidence for the "one gene-one enzyme" hypothesis, using mutants of Neurospora crassa deficient in a specific enzyme called glutamate dehydrogenase. Fincham was appointed first as lecturer in botany (1950-1954) and then as reader (1954-1960) at University of Leicester. A year as an associate professor in the Massachusetts Institute of Technology preceded his appointment as head ...
Suggest two methods to isolate a collection of cold-sensitive mutants in Salmonella typhimurium -- that is, mutants that grow at 42 C but do not grow at 30 C.. ANSWER: Although conditional mutations may not grow at the nonpermissive temperature, they often survive short exposure to the nonpermissive temperature. With this hint, consider the three basic approaches for isolating mutants: selections, screens, and enrichments. There is no obvious way of selecting for the desired mutants because the desired mutation is unable to grow under the nonpermissive conditions. It would be straightforward to screen for the desired mutants -- for example, (i) you could plate colonies at 30 C, replica plate the colonies to 30 C and 42 C, then look for colonies that grow at 42 C but not at 30 C, or (ii) you could plate the cells at the nonpermissive temperature for a short time then shift the plates to the permissive temperature -- the mutants usually form smaller colonies due to the effect of the temporary ...
Linkage of at least two complementation groups of ataxia-telangiectasia (AT) to the chromosomal region 11q23 is now well established. We provide here an 18-point map of the surrounding genomic region, derived from linkage analysis of 40 CEPH families. On the basis of this map, 111 AT families from Turkey, Israel, England, Italy, and the United States were analyzed, localizing the AT gene(s) to an 8-cM sex-averaged interval between the markers STMY and D11S132/NCAM. A new Monte Carlo method for computing approximate location scores estimates this location as being at least 10(8) times more likely than the next most likely interval, with a support interval midway between STMY and D11S132 that is either 5 ...
Haider, Ameena J. and Briggs, Deborah and Self, Tim J. and Chilvers, Hannah L. and Holliday, Nicholas D. and Kerr, Ian D. (2011) Dimerization of ABCG2 analysed by bimolecular fluorescence complementation. PLoS ONE, 6 (10). e25818/1-e25818/9. ISSN 1932-6203 ...
My research focuses on design of peptide antagonists with high affinity and specificity for inhibiting protein-protein interactions. While forces driving stability are now well understood, much less is known about designing specificity. We use a Protein-fragment Complementation Assay (PCA), which utilises semi-rational design to select antagonists of Jun and Fos peptides that interact in the oncogenic Activator Protein-1 transcriptional regulator. We have devised a Competitive and Negative Design Initiative (CANDI) technique to increase target-specificity in the PCA system by expressing potential off-targets during selection. Peptides generated have allowed us to predict coiled coil stability and specificity based only the primary sequence, and to consequently design peptides. Our screening approach is also applied to a range of other therapeutically relevant systems that include β-amyloid and α-synuclein implicated in the pathogenesis of Alzheimers and Parkinsons diseases, respectively. ...
Cassonnet P, Rolloy C, Neveu G, Vidalain P-O, Chantier T, Pellet J, Jones L, Muller M, Demeret C, Gaud G et al.. 2011. Benchmarking a luciferase complementation assay for detecting protein complexes. Nature Methods. 8:990-992. ...
However, in reality, no two segments of a polymer chain can occupy the same point in space. This means, instead of being described by the random walk, it is described by the self-avoiding random walk. Because the self-avoiding walk excludes configurations that visit the same site, it is generally bigger than a self-avoiding walk of the same length: if you "turn on" self-avoidance interactions, the chain "swells" so this is called a swollen chain. So, if the ideal chain grows as ##L^{0.5}##, how does the swollen chain grow with length?. If we consider some number N particles each with volume v inside a larger volume V, the probability of any two particles occupying the same space is. $$p_{1}=\frac{v}{V}$$. So the probability of them not occupying the same space is:. $$\overline{\,p_{1}}=1-\frac{v}{V}$$. Now, if there are N of these particles, there are N(N-1)/2 pairwise combinations of them, so the probability of no two particles occupying the same space ...
Book now at Benihana - Dulles in Dulles, explore menu, see photos and read 480 reviews: Its Benihana, so how bad can it be? This one tries to answer the question.
Good skin care products do not only make your skin look young but they also should keep your skin healthy. In order to do this these products need to contain natural ingredients in the right proportion. This way they can complement each other and work together.|br/|The products should...
If one parent is a wild-type mouse and the other is a homozygous knockout mouse, their offspring will be heterozygous at the knockout gene. The mouse will likely produce the protein from the wild-type copy of the gene, but depending on how the gene is regulated it is likely that expression of the protein will be below wild-type levels ...
Human peroxisome biogenesis disorders are lethal genetic diseases in which abnormal peroxisome assembly compromises overall peroxisome and cellular function. Peroxisomes are ubiquitous membrane-bound organelles involved in several important biochemical processes, notably lipid metabolism and the use of reactive oxygen species for detoxification. Using cultured cells, we systematically characterized the peroxisome assembly phenotypes associated with dsRNA-mediated knockdown of 14 predicted Drosophila homologs of PEX genes (encoding peroxins; required for peroxisome assembly and linked to peroxisome biogenesis disorders), and confirmed that at least 13 of them are required for normal peroxisome assembly. We also demonstrate the relevance of Drosophila as a genetic model for the early developmental defects associated with the human peroxisome biogenesis disorders. Mutation of the PEX1 gene is the most common cause of peroxisome biogenesis disorders and is one of the causes of the most severe form ...

Method for treating a metastatic carcinoma using a conditionally lethal     gene - Patent # 5997859 - PatentGeniusMethod for treating a metastatic carcinoma using a conditionally lethal gene - Patent # 5997859 - PatentGenius

In order to test whether in vivo transduction of a murine tumor could be used to treat the disease, an experiment was performed ... Of these, extensive genetic and biochemical characterization have been performed on Ad2 and Ad5. Id. at 618. Based upon these ... to deletions which can be made in the dispensable E3 region and in E1using complementation on 293 cells, and to the fact that ... 18). These vectors are used as pseudo-HIV to test-activate tat-dependent HSVTK vectors.. 1. The His.sup.r expression vector ...
more infohttp://www.patentgenius.com/patent/5997859.html

Frontiers | Analysis of Magnaporthe oryzae Genome Reveals a Fungal Effector, Which Is Able to Induce Resistance Response in...Frontiers | Analysis of Magnaporthe oryzae Genome Reveals a Fungal Effector, Which Is Able to Induce Resistance Response in...

Genetic complementation test also proved that Mo-01947_9 protein induces avirulence response in the pathogen in the presence of ... Genetic complementation test also proved that Mo-01947_9 protein induces avirulence response in the pathogen in the presence of ... Genetic Complementation Test of Candidate AvrPi54 Gene. Complementation of avirulence function of the cloned candidate AvrPi54 ... Genetic complementation test also proved that Mo-01947_9 protein induces avirulence response in the pathogen in presence of ...
more infohttps://www.frontiersin.org/articles/10.3389/fpls.2016.01140/full

Molecular cloning and functional analysis of a blue light receptor gene MdCRY2 from apple (Malus domestica).Molecular cloning and functional analysis of a blue light receptor gene MdCRY2 from apple (Malus domestica).

Genetic Complementation Test. Light. Malus / genetics*, growth & development, metabolism. Molecular Sequence Data. Plant ... the interaction between MdCRY2 and AtCOP1 using a yeast two-hybrid assay and a bimolecular fluorescence complementation assay. ...
more infohttp://www.biomedsearch.com/nih/Molecular-cloning-functional-analysis-blue/23314496.html

Complementation in Zellweger syndrome: biochemical analysis of newly generated peroxisomes.Complementation in Zellweger syndrome: biochemical analysis of newly generated peroxisomes.

Genetic Complementation Test. Humans. Microbodies / metabolism*, ultrastructure. Oxidation-Reduction. Zellweger Syndrome / ... Since this disorder is genetically heterogeneous and several complementation groups are known, we were able to study the ... normalization of peroxisomal activity during the process of complementation. The restoration of catalase and dihydroxyacetone ...
more infohttp://www.biomedsearch.com/nih/Complementation-in-Zellweger-syndrome-biochemical/1511996.html

Complementation of Yeast Genes with Human Genes as an Experimental Platform for Functional Testing of Human Genetic Variants |...Complementation of Yeast Genes with Human Genes as an Experimental Platform for Functional Testing of Human Genetic Variants |...

Complementation of Yeast Genes with Human Genes as an Experimental Platform for Functional Testing of Human Genetic Variants. ... Complementation of Yeast Genes with Human Genes as an Experimental Platform for Functional Testing of Human Genetic Variants. ... Complementation of Yeast Genes with Human Genes as an Experimental Platform for Functional Testing of Human Genetic Variants. ... Complementation of Yeast Genes with Human Genes as an Experimental Platform for Functional Testing of Human Genetic Variants ...
more infohttp://www.genetics.org/content/201/3/1263

MeSH ORA framework: R/Bioconductor packages to support MeSH over-representation analysis | BMC Bioinformatics | Full TextMeSH ORA framework: R/Bioconductor packages to support MeSH over-representation analysis | BMC Bioinformatics | Full Text

Genetic Complementation Test (+8). Promoter Regions, Genetic (+24). PA5471. Ribosomes. Pseudomonas aeruginosa ... 2) Multiplicity of tests * As described above, in ORA, several thousand statistical tests are conducted simultaneously. Such an ... The hypergeometric test (or Fishers exact test) is widely used to calculate such probabilities. Several thousand statistical ... Genetic Epidemiol. 2002; 23:70-86.View ArticleGoogle Scholar. *. Durinck S, Spellman PT, Birney E, Huber W. Mapping identifiers ...
more infohttps://bmcbioinformatics.biomedcentral.com/articles/10.1186/s12859-015-0453-z

Smyd3 methylates H4K5 in vitro and in vivo. (A) Methyla | Open-iSmyd3 methylates H4K5 in vitro and in vivo. (A) Methyla | Open-i

Genetic Complementation Test. *HeLa Cells. *Humans. *Methylation. *Mice. *Mice, Inbred C57BL. *Mice, Knockout ...
more infohttps://openi.nlm.nih.gov/detailedresult.php?img=PMC3368817_epi-7-340-g2&req=4

A Multifaceted Genomics Approach Allows the Isolation of the Rice Pia-blast Resistance Gene Consisting of Two Adjacent NBS-LRR...A Multifaceted Genomics Approach Allows the Isolation of the Rice Pia-blast Resistance Gene Consisting of Two Adjacent NBS-LRR...

Genetic Complementation Test Actions. * Search in PubMed * Search in MeSH * Add to Search ... An additional complementation study revealed that the two NBS-LRR-type R genes, SasRGA4 and SasRGA5, that are located next to ... High-resolution genetic mapping and sequencing of the gene region in the Pikm-containing c … ...
more infohttps://pubmed.ncbi.nlm.nih.gov/21251109/

The CG12120 Gene Product Possesses NBAD Hydrolase and N | Open-iThe CG12120 Gene Product Possesses NBAD Hydrolase and N | Open-i

Genetic Complementation Test. *Molecular Sequence Data. *Mutation. *Sequence Homology, Amino Acid. Related in: MedlinePlus ... To test whether CG12120 possesses these predicted activities, we produced recombinant CG12120 protein using a baculovirus ... To test whether CG12120 possesses these predicted activities, we produced recombinant CG12120 protein using a baculovirus ... This is, to our knowledge, the first molecular genetic characterization of NBAD hydrolase and carcinine hydrolase activity in ...
more infohttps://openi.nlm.nih.gov/detailedresult.php?img=PMC1285064_pgen.0010063.g004&req=4

Beta alpha)8-barrel proteins of tryptophan biosynthesis in the hyperthermophile Thermotoga maritima.  - University of...Beta alpha)8-barrel proteins of tryptophan biosynthesis in the hyperthermophile Thermotoga maritima. - University of...

Genetic Complementation Test. MESH. Gram-Negative Anaerobic Bacteria/genetics. MESH. Hot Temperature. MESH. ... The genes were cloned by complementation in vivo of trp deletion strains of Escherichia coli. The new sequences, together with ... The genes were cloned by complementation in vivo of trp deletion strains of Escherichia coli. The new sequences, together with ...
more infohttps://epub.uni-regensburg.de/13715/

Five components of the ethylene-response pathway identified in a screen for weak ethylene-insensitive mutants in Arabidopsis |...Five components of the ethylene-response pathway identified in a screen for weak ethylene-insensitive mutants in Arabidopsis |...

Genetic complementation testing indicated that wei2 and wei3 were not alleles of the EIN2 gene. The wei2 mutant was further ... 6). Genetic complementation analysis indicated that wei1 and tir1-1 were allelic (data not shown). Sequencing of the TIR1 gene ... Genetic and Phenotypic Characterization.. Genetic analysis of the progeny produced from backcrosses of these five mutants to ... To test this hypothesis, we crossed the eil1 mutants to ein3-1. The F1 progeny of the crosses between eil1-1 or eil1-2 and ein3 ...
more infohttps://www.pnas.org/content/100/5/2992?ijkey=98cd93f393897c5cf7d3027f17adacdfc0e712f5&keytype2=tf_ipsecsha

ZFIN Publication: Abdelilah et al., 1996ZFIN Publication: Abdelilah et al., 1996

Genetic Complementation Test. *Mutation*. *Nerve Degeneration/genetics. *Neurons/physiology*. *Phenotype. *Rhombencephalon/ ... To prevent accidental cell death, mechanisms that trigger programmed cell death, as well as the genetic components of the cell ... The mutations presented here might provide a genetic framework to aid in the understanding of the etiology of degenerative and ...
more infohttp://zfin.org/ZDB-PUB-970210-18

ZFIN Publication: Granato et al., 1996ZFIN Publication: Granato et al., 1996

... we included a simple touch response test in our zebrafish large-scale genetic screen. In total we identified 166 mutants with ... Genetic Complementation Test. *Larva/physiology. *Locomotion/genetics*. *Muscles/embryology. *Mutation*. *Phenotype. *Somites/ ...
more infohttps://zfin.org/ZDB-PUB-970210-34

Max Planck Society - eDoc ServerMax Planck Society - eDoc Server

Genetic Complementation Test/methods; Genetic Loci; Green Fluorescent Proteins/metabolism; Immunoprecipitation/methods; MADS ... Despite extensive genetic studies, little is known about the transcriptional control of SOC1, and we are just starting to ... Domain Proteins/genetics/*metabolism; Promoter Regions, Genetic; Protein Binding; *Regulatory Sequences, Nucleic Acid; Signal ...
more infohttp://edoc.mpg.de/display.epl?mode=doc&id=636050&col=65&grp=326

Welcome to LibAge, the ageing reference resourceWelcome to LibAge, the ageing reference resource

Entries tagged Genetic Complementation Test (98). Sort:. Number of citations. Author. Title. Year. PubMed ID. ... A curated database of genes associated with dietary restriction in model organisms either from genetic manipulation experiments ...
more infohttp://la.ageing-map.org/entries/tags/193/

Welcome to LibAge, the ageing reference resourceWelcome to LibAge, the ageing reference resource

Entries tagged Genetic Complementation Test (98). Sort:. Number of citations. Author. Title. Year. PubMed ID. ... A curated database of genes associated with dietary restriction in model organisms either from genetic manipulation experiments ...
more infohttp://libage.ageing-map.org/entries/tags/193/

A nuclear magnetic resonance based approach to accurate functional annotation of putative enzymes in the methanogen...A nuclear magnetic resonance based approach to accurate functional annotation of putative enzymes in the methanogen...

For a number of case studies, additional experiments (such as in vivo genetic complementation) were performed to determine ... Here, we describe the use of nuclear magnetic resonance (NMR)-based ligand screening as a tool for testing functional ... The functional annotation of MA4265 was further tested using genetic complementation studies. The E. coli genome contains three ... Gene complementation. Genetic complementation of E. coli mutants by the MA genes was performed with E. coli deletion mutants ...
more infohttps://bmcgenomics.biomedcentral.com/articles/10.1186/1471-2164-12-S1-S7

Ric-8 Proteins Are Molecular Chaperones That Direct Nascent G Protein α Subunit Membrane Association | Science SignalingRic-8 Proteins Are Molecular Chaperones That Direct Nascent G Protein α Subunit Membrane Association | Science Signaling

We then performed genetic complementation tests of the Ric-8A- or Ric-8B-null cell defects. A Ric-8A−/− mES cell line made to ... We tested the effects of complete ablation of Ric-8A or Ric-8B on steady-state G protein amounts in crude membrane fractions ( ... A genetic selection for Caenorhabditis elegans synaptic transmission mutants. Proc. Natl. Acad. Sci. U.S.A. 93, 12593-12598 ( ... Gα and Gβγ subunit abundances are interdependent (33). We tested whether transient transfection of Ric-8A−/− and Ric-8B−/− mES ...
more infohttps://stke.sciencemag.org/content/4/200/ra79?ijkey=9b55a0d8b3f1bb79100144d5fa6ac96c0531e450&keytype2=tf_ipsecsha

Effects of mutating α-tubulin lysine 40 on sensory dendrite development | Journal of Cell ScienceEffects of mutating α-tubulin lysine 40 on sensory dendrite development | Journal of Cell Science

Genetic complementation tests also unexpectedly revealed that αTub84D is a non-essential gene (Table 1). Combined, these data ... 1A). We tested whether the function of the α-tubulin C-terminal tails from flies and mammals might be conserved despite these ... Two-tailed Students t-tests were used to compare two conditions. *P=0.05-0.01; **P=0.01-0.001; ***P=0.0001-0.001; ****P,0.0001 ... To test the role of αTub84B K40 acetylation in survival and neuronal morphogenesis, we introduced K40A and K40R mutations to ...
more infohttp://jcs.biologists.org/content/130/24/4120?utm_source=INT&utm_medium=FP&utm_campaign=JCS_Snippet&utm_content=130-4120

Related Arabidopsis Serine Carboxypeptidase-Like Sinapoylglucose Acyltransferases Display Distinct But Overlapping Substrate...Related Arabidopsis Serine Carboxypeptidase-Like Sinapoylglucose Acyltransferases Display Distinct But Overlapping Substrate...

... and their biochemical sng1 phenotypes were confirmed by HPLC and genetic complementation tests. The genomic regions deleted ... To test this hypothesis, extracts from a variety of tissues from the sng1-5 and sng1-6 mutants were analyzed via HPLC to ... To test the hypothesis that At2g23010 is required for the synthesis of compound 1 and 1,2-disinapoyl-Glc, the sng1-6 mutant was ... To test the hypothesis that one or more of the SCPL genes clustered near At2g23010 also have SST activity, two additional sng1- ...
more infohttp://www.plantphysiol.org/node/26385.full.print

Mary E Porter - Research Output
     - Experts@MinnesotaMary E Porter - Research Output - [email protected]

Reappraisal of the genetic map of Chlamydomonas reinhardtii. Dutcher, S. K., Power, J., Galloway, R. E. & Porter, M. E., Jan 1 ...
more infohttps://experts.umn.edu/en/persons/mary-e-porter/publications/?ordering=publicationYearThenTitle&descending=false

hyp Loci Control Cell Pattern Formation in the Vegetative Mycelium of Aspergillus nidulans | Geneticshyp Loci Control Cell Pattern Formation in the Vegetative Mycelium of Aspergillus nidulans | Genetics

... and used to test for genetic complementation. Diploids made from crosses between hyp and wild-type strains were used to test ... To test if the loss of hypA was capable of overriding nuclear cycle blocks, we constructed double mutants containing hypA1 and ... To test this hypothesis, average subapical cell volumes were calculated using the data in Table 3, A and C. Average cell ... 1994 Genetic requirements for initiating asexual development in Aspergillus nidulans. Curr. Genet. 27: 62-69. ...
more infohttps://www.genetics.org/content/148/2/669?ijkey=3985f69e3076062d785072d359c4cd56cb72c4f5&keytype2=tf_ipsecsha

000646 - A/J000646 - A/J

Our preclinical efficacy testing services offer scientific expertise and an array of target-based and phenotype-based outcome ... p,A/J inbred mice are widely used to model cancer and for carcinogen testing given their high susceptibility to carcinogen- ... Genetic complementation tests have shown allelism between the mdfw (modifier of deaf waddler) locus and the ahl locus. Further ... In addition, a standard genetic test of allelism between clf1 and a Wnt9b targeted mutation demonstrated noncomplementation, ...
more infohttps://www.jax.org/strain/000646

000653 - BUB/BnJ000653 - BUB/BnJ

Genetic complementation tests have shown allelism between the mdfw (modifier of deaf waddler) locus and the ahl locus. Further ... The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic ... Genetic influence on electrocardiogram time intervals and heart rate in aging mice. Am J Physiol Heart Circ Physiol 296(6): ... Identification of genetic determinants of IGF-1 levels and longevity among mouse inbred strains. Aging Cell 9(5):823-36PubMed: ...
more infohttps://www.jax.org/strain/000653
  • To test whether a methylation-sensitive restriction system contributes to poor B. burgdorferi transformability, shuttle plasmids were treated with the CpG methylase M.SssI prior to the electroporation of a variety of strains harboring different putative R-M systems. (umassmed.edu)
  • RIC-8 was discovered during a genetic screen of Caenorhabditis elegans that was designed to uncover mutants with defective neurotransmitter release ( 3 ). (sciencemag.org)
  • A/J inbred mice are widely used to model cancer and for carcinogen testing given their high susceptibility to carcinogen-induced tumors. (jax.org)
  • To test whether CG12120 possesses these predicted activities, we produced recombinant CG12120 protein using a baculovirus expression system in insect cell culture (Figure 4). (nih.gov)
  • The nematode Caenorhabditis elegans has provided a powerful system for dissecting the genetic mechanisms controlling animals' intrinsic biology and interaction with environmental factors. (g3journal.org)
  • This is, to our knowledge, the first molecular genetic characterization of NBAD hydrolase and carcinine hydrolase activity in any organism and is central to the understanding of pigmentation and photoreceptor function. (nih.gov)