Philip M. Service The author is in the Department of Biological Sciences, Northern Arizona University, Flagstaff, AZ 86011, USA. E-mail: Philip.Service{at}nau.edu. http://sageke.sciencemag.org/cgi/content/full/2004/12/pe13 Key Words: quantitative complementation test life-span gene Drosophila quantitative genetics QTL mapping. Abstract: Several recent studies have used quantitative complementation tests to identify relatively short chromosome regions that contain genes that influence life span and to screen for candidate life-span genes in flies. The methodology and logic of quantitative complementation tests are described. Arguments are presented that suggest that these tests may be misleading because there is a substantial, but unknown, likelihood of false positive results. The arguments are supported by the published results of quantitative complementation tests.. Citation: P. M. Service, How Good Are Quantitative Complementation Tests? Sci. Aging Knowl. Environ. 2004 (12), pe13 (2004). ...
TY - JOUR. T1 - Recovery of YAC-end sequences through complementation of an Escherichia coli pyrF mutation. AU - Wright, David A.. AU - Park, Sei Kyoung. AU - Wu, Dongying. AU - Phillips, Gregory J.. AU - Rodermel, Steven R.. AU - Voytas, Daniel F.. PY - 1997/7/1. Y1 - 1997/7/1. N2 - We have developed a genetic means to recover sequences from YAC-ends near the yeast selectable marker URA3. This strategy is based on the ability of URA3 to complement mutations in pyrF, an Escherichia coli gene required for pyrimidine biosynthesis. We have developed an E. coli strain with a non-reverting allele of pyrF that is also suitable for cloning (recA-, hsdR-). We demonstrate the utility of this complementation strategy to obtain right-end clones from three YACs containing Arabidopsis thaliana DNA.. AB - We have developed a genetic means to recover sequences from YAC-ends near the yeast selectable marker URA3. This strategy is based on the ability of URA3 to complement mutations in pyrF, an Escherichia coli ...
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. ...
The development of high-throughput and large-scale technologies have expanded the screening capacity for human-yeast complementation pairs. As a result, several systematic screens have reported testing the essential yeast genes for replaceability (Zhang et al. 2003; Hamza et al. 2015; Kachroo et al. 2015; Sun et al. 2016; Yang et al. 2017; Garge et al. 2019; Laurent et al. 2019). These studies generated overlapping lists of human-yeast complementation pairs and arrived at similar conclusions regarding features that predict the replaceability of essential yeast genes. However, compared to the essential yeast genes, the nonessential genes are a much larger set and have a variety of different phenotypic readouts, making them more difficult to screen systematically for complementation. In this study, we have started this process by focusing on a subset of the nonessential yeast genes, specifically those required for chromosome maintenance. We identified 20 complementation pairs that are replaceable ...
Occurs when wild type phenotype is restored in an F1 individual made by crossing two independent mutants, carrying different heteroalleles
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.
TY - JOUR. T1 - In vitro complementation as an assay for purification of adenovirus DNA replication proteins. AU - Ostrove, J. M.. AU - Rosenfeld, P.. AU - Williams, J.. AU - Kelly, T.. PY - 1983. Y1 - 1983. N2 - As an approach to the purification of adenovirusencoded DNA replication proteins, we have developed in vitro complementation assays that make use of viral mutants defective in DNA replication in vivo. Nuclear extracts prepared from cells infected with H5ts36 or H5ts125, two such mutants belonging to different complementation groups, were found to be defective in viral DNA replication in vitro. However, replication activity could be restored by mixing the two extracts. Replication activity in either extract also could be restored by addition of appropriate replication-deficient fractions purified from cells infected with wild-type adenovirus. By using such assays, H5ts36- and H5ts125-complementing activities were extensively purified. As expected, purified H5ts125-complementing activity ...
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 ...
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 ...
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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
2001b, 2007, 2008). Mutation in the lytM gene was subsequently transduced into the S. aureus lyt− strain (Mani et al., 1993; Ramadurai & Jayaswal, 1997) to potentially create an autolysin-free lyt−:lytM double mutant. For genetic complementation of the lytM mutant, an approximately 2.2-kb DNA fragment was PCR amplified using primers P5 and P6 and S. aureus SH1000 genomic DNA as template. This amplicon represents a fragment starting 890 nt upstream and ending 364 nt downstream of the lytM gene that was cloned into the BamHI and HindIII Screening Library manufacturer sites. of shuttle plasmid pCU1 (Augustin et al., 1992) and subsequently transferred to a lytM mutant of S. aureus SH1000. Mid-exponential-phase cultures (OD600 nm=0.6) were diluted 50-fold in a nephelo culture flask (Wheaton) containing 50 mL fresh TSB with a flask-to-medium volume ratio of 6 : 1 and growth was followed by measurement of OD600 nm spectrophotometrically. In another experiment, cultures pregrown to an OD600 nm=0.5 ...
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.
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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.
TY - JOUR. T1 - Genetic diversity of UV-sensitive DNA repair mutants of Chinese hamster ovary cells. AU - Thompson, L. H.. AU - Busch, D. B.. AU - Brookman, K.. AU - Mooney, C. L.. AU - Glaser, D. A.. PY - 1981. Y1 - 1981. N2 - Mutant lines of Chinese hamster ovary cells that show hypersensitivity to killing and mutagenesis by UV light were analyzed by genetic complementation analysis to determine whether defects in different gene loci might underlie a common cellular phenotype. To facilitate rapid screening of mutant clones, a procedure was devised that allowed presumptive complementation to be assessed on the basis of the frequency of UV-resistant cells after fusion by polyethylene glycol. Four classes were identified among 44 clones tested. By using drug-resistance markers for selection of hybrid cells in crosses between UV mutant and wild type, a mutant from each of the four classes was shown to behave as phenotypically recessive. Hybrids were also isolated from crosses between each of the ...
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 ...
Xeroderma pigmentosum (XP) is a rare recessive disorder that is characterized by extreme sensitivity to UV light. UV light exposure results in the formation of DNA damage such as cyclobutane dimers and (6-4) photoproducts. Nucleotide excision repair (NER) orchestrates the removal of cyclobutane dimers and (6-4) photoproducts as well as some forms of bulky chemical DNA adducts. The disease XP is comprised of 7 complementation groups (XP-A to XP-G), which represent functional deficiencies in seven different genes, all of which are believed to be involved in NER. The main clinical feature of XP is various forms of skin cancers; however, neurological degeneration is present in XPA, XPB, XPD and XPG complementation groups. The relationship between NER and other types of DNA repair processes is now becoming evident but the exact relationships between the different complementation groups remains to be precisely determined. Using gene expression analysis we have identified similarities and differences after UV
DNA synthesis in vitro in Brij-treated Saccharomyces cerevisiae requires the product of the CDC8 gene (Hereford, L. M. & Hartwell, L. H. (1971) Nature (London) New Biol. 234, 171-172). Extracts of wild-type A364a yeast restore DNA synthesis in Brij-treated cdc8, a mutant containing a thermolabile cdc8 gene product. This constitutes a complementation assay by which the cdc8 gene product can be monitored during purification. A heat-stable protein responsible for this complementation has been partially purified from both wild-type A364a cells and from a cdc8 temperature-sensitive mutant. The complementation activity from the mutant is thermolabile when compared to the wild-type activity, indicating that CDC8 is the structural gene for the protein. ...
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 ...
For a simple example of a complementation test, suppose a geneticist is interested in studying two strains of white-eyed flies of the species Drosophila melanogaster, more commonly known as the common fruit fly. In this species, wild type flies have red eyes and eye color is known to be related to two genes, A and B. Each one of these genes has two alleles, a dominant one that codes for a working protein (A and B respectively) and a recessive one that codes for a malfunctioning protein (a and b respectively). Since both proteins are necessary for the synthesis of red pigmentation in the eyes, if a given fly is homozygous for either a or b, it will have white eyes. Knowing this, the geneticist may perform a complementation test on two separately obtained strains of pure-breeding white-eyed flies. The test is performed by crossing two flies, one from each strain. If the resulting progeny have red eyes, the two strains are said to complement; if the progeny have white eyes, they do not. If the ...
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 ...
We report here, to our knowledge, the first biochemical characterization of a plant biotin synthase reaction. Heterologous interactions between a plant recombinant Bio2 protein and bacterial proteins yield a functional biotin synthase complex, in good agreement with the successful functional complementation approach, using anE. coli bioB mutant, employed to isolate the bio2gene product from Arabidopsis (Baldet and Ruffet, 1996). The turnover number of the reaction was ,2 h−1 in the heterologous system with unfractionated protein extract from Bio2-overproducing strain and still ,1 h−1in the heterologous system comprising purified Bio2 protein (calculated from data in Table I). It appears from our results that biotin synthase from Arabidopsis acts as a catalyst and not, as suggested in bacteria, as a simple reactant (Gibson et al., 1999; Kiyasu et al., 2000).. The relative low levels of biotin synthase measured in our in vitro systems may reflect the limited proportion of recombinant Bio2 ...
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 ...
A supplementary New Mutants Annual series began in 1984. These annuals were always written by whoever was the regular New Mutants writer at the time and often included significant changes to the status quo which were not explained in the parent series so that readers would have to buy New Mutants Annual in order to follow events in both series.1985 annual was solicited as New Mutants Annual #2, but published as New Mutants Special Edition #1 because it exceeded the maximum page count for an annual.[5] In 1986, Professor X was written out of the series. Before he left, he made the X-Mens one-time nemesis, Magneto, headmaster of his school.[6] Magneto would be the teams longest-running headmaster, holding the position from New Mutants #35 through to #75. Fiercely overprotective of his students, particularly after the events of the Mutant Massacre and Fall of the Mutants, he was increasingly used as an uptight foil for the adventurous New Mutants, setting rules that they would inevitably ...
Cellular biochemical machineries, what we call pathways, consist of dynamically assembling and disassembling macromolecular complexes. Although our models for the organization of biochemical machines...
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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.
TY - JOUR. T1 - Characterization of a complex chromosomal rearrangement maps the locus for In vitro complementation of xeroderma pigmentosum group D to human chromosome band 19q 13. AU - Flejter, W. L.. AU - McDaniel, L. D.. AU - Askari, M.. AU - Friedberg, E. C.. AU - Schultz, R. A.. N1 - Copyright: Copyright 2016 Elsevier B.V., All rights reserved.. PY - 1992/11. Y1 - 1992/11. N2 - Microcell‐mediated chromosome transfer (MMCT) is a powerful genetic technique that permits the transfer of a single chromosome from one mammalian cell to another. The utility of MMCT for gene mapping strategies is critically dependent on the careful characterization of the chromosomes being transferred. We have recently reported the identification of a single rearranged human chromosome, designated Tneo, which corrects the UV sensitivity and excision repair defect of cells of xeroderma pigmentosum genetic complementation group D (XP‐D) in culture (Flejter WL et al., Proc Natl Acad Sci USA 89:261-265, 1992). ...
TY - JOUR. T1 - The peroxin Pex6p gene is impaired in peroxisomal biogenesis disorders of complementation group 6. AU - Matsumoto, N.. AU - Tamura, S.. AU - Moser, Ann B.. AU - Moser, H. W.. AU - Braverman, N.. AU - Suzuki, Y.. AU - Shimozawa, N.. AU - Kondo, N.. AU - Fujiki, Y.. PY - 2001. Y1 - 2001. N2 - Human genetic peroxisomal biogenesis disorders (PBDs), such as Zellweger syndrome, comprise 13 different complementation groups (CGs). Eleven peroxin genes, termed PEXs, responsible for PBDs have been identified, whereas pathogenic genes for PBDs of 2CGs, CG-A (the same CG as CG8 in the United States and Europe) and CG6, remained unidentified. We herein provide several lines of novel evidence indicating that PEX6, the pathogenic gene for CG4, is impaired in PBD of CG6. Expression of PEX6 restored peroxisome assembly in fibroblasts from a CG6 PBD patient. This patient was a compound heterozygote for PEX6 gene alleles. Accordingly, by merging CG6 with CG4, human PBDs are now classified into 12 ...
To further our studies of protein sorting and biogenesis of the lysosome-like vacuole in yeast, we have isolated spontaneous mutations in 11 new VPL complementation groups, as well as additional alleles of the eight previously described VPL genes. These mutants were identified by selecting for cells that mislocalize vacuolar proteins to the cell surface. Morphological examination of the vpl mutants indicated that most contain vacuoles of normal appearance; however, some of the mutants generally lack a large vacuole, and instead accumulate smaller organelles. Of the 19 VPL complementation groups, 12 were found to be identical to 12 of 33 VPT complementation groups identified in a separate study. Moreover, the end1 mutant and all of the previously reported pep mutants, with the exception of pep4, were found to exhibit a profound vacuolar protein sorting defect, and complementation tests between the PEP, VPL VPT and END1 groups demonstrated that there are extensive overlaps between these groups. ...
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
A split-EGFP bimolecular fluorescence complementation assay was used to visualise and locate three interacting pairs of proteins from the GAL genetic switch of the budding yeast, Saccharomyces cerevisiae. Both the Gal4p-Gal80p and Gal80p-Gal3p pairs were found to be located in the nucleus under indu …
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
Montibello - Spanish hair care philosophy bases on complementation of essential nourishing ingredients. Treat Naturtech product line is a 100% realisation of
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 ...
Background Hereditary evidence in indicates that members from the Snf1-Related Kinases 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.
Characterization of a Regulator pgsR on Endogenous Plasmid p2Sip and Its Complementation for Poly(γ-glutamic acid) Accumulation in Bacillus amyloliquefaciens
Eurofins DiscoverX has created a series of cell-based assays for biologics and small molecule applications based on our robust Enzyme Fragment Complementation (EFC) platform. This webinar series focuses on unique applications for protein degradation, signaling pathways, and more, and explores EFC cell-based assays in combination with PROTACs, CRISPR, biosensor development, and reporter technologies ...