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URA3 is often used in yeast research as a "marker gene", that is, a gene to label chromosomes or plasmids. URA3 encodes Orotidine 5'-phosphate decarboxylase (ODCase) , which is an enzyme that catalyzes one reaction in the synthesis of pyrimidine ribonucleotides (a component of RNA ).
Figure 2: The bacteria one-hybrid system requires two customized plasmids: (a) the “bait” vector (pB1H1 or pB1H2) which expresses the DNA-binding domain as a fusion construct with a subunit (α or ω) of RNA polymerase, and (b) a reporter plasmid (pH3U3) containing selectable markers and a binding-site library, or “prey,” which will potentially interact with the "bait.”
In yeast and bacteria, OMP decarboxylase is a single-function enzyme.However, in mammals, OMP decarboxylase is part of a single protein with two catalytic activities.This bifunctional enzyme is named UMP synthase and it also catalyzes the preceding reaction in pyrimidine nucleotide biosynthesis, the transfer of ribose 5-phosphate from 5-phosphoribosyl-1-pyrophosphate to orotate to form OMP.
5-Fluoroorotic acid (5FOA) is a fluorinated derivative of the pyrimidine precursor orotic acid.It is used in yeast genetics to select for the absence of the URA3 gene, which encodes the enzyme for the decarboxylation of 5-fluoroorotic acid to 5-fluorouracil, a toxic metabolite. [1]
The yeast deletion project, formally the Saccharomyces Genome Deletion Project, is a project to create data for a near-complete collection of gene-deletion mutants of the yeast Saccharomyces cerevisiae. Each strain carries a precise deletion of one of the genes in the genome. This allows researchers to determine what each gene does by comparing ...
Shuttle vectors are frequently used to quickly make multiple copies of the gene in E. coli (amplification). They can also be used for in vitro experiments and modifications (e.g. mutagenesis, PCR). One of the most common types of shuttle vectors is the yeast shuttle vector. [2] Almost all commonly used S. cerevisiae vectors are shuttle vectors.
A conditional gene knockout allows gene deletion in a tissue in a tissue specific manner. This is required in place of a gene knockout if the null mutation would lead to embryonic death, [13] or a specific tissue or cell type is of specific interest. This is done by introducing short sequences called loxP sites around the gene.
Y187 is a strain of yeast (Saccharomyces cerevisiae) used in biological research for two-hybrid screening.The strain has been sold commercially by Clontech [1] since at least 2000 and is used as a partner with strain AH109 in mating assays.