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.