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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 ).
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]
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.”
Neo gene from Tn5, which confers resistance to kanamycin in bacteria and geneticin in eukaryotic cells. [3] Mutant FabI gene (mFabI) from the E. coli genome, which confers triclosan resistance to the host. [4] URA3, an orotidine-5' phosphate decarboxylase from yeast, is a positive and negative selectable marker. It is required for uracil ...
Auxotrophic selection markers that allow an auxotrophic organism to grow in minimal growth medium may also be used; examples of these are LEU2 and URA3 which are used with their corresponding auxotrophic strains of yeast. [7] Another kind of selectable marker allows for the positive selection of plasmid with cloned gene.
The URA3 marker (URA3 blaster method) is an often-used strategy in uridine auxotrophic strains; however, studies have shown that differences in URA3 position in the genome can be involved in the pathogeny of C. albicans. [119] Besides the URA3 selection one can also use the histidine, leucine and arginine autotrophy.
For example, the notation "YEL021W", an alias of the URA3 gene used in the National Center for Biotechnology Information (NCBI) database, denotes that this gene is in the 21st open reading frame (ORF) from the centromere of the left arm (L) of Yeast (Y) chromosome number V (E), and that the expression coding strand is the Watson strand (W ...