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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 synthesis pathway gene is a uracil auxotroph (e.g., if the yeast Orotidine 5'-phosphate decarboxylase gene is inactivated, the resultant strain is a uracil ...
Two-hybrid screening (originally known as yeast two-hybrid system or Y2H) is a molecular biology technique used to discover protein–protein interactions (PPIs) [1] and protein–DNA interactions [2] [3] by testing for physical interactions (such as binding) between two proteins or a single protein and a DNA molecule, respectively.
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 ).
Next to the above-mentioned selection makers a few auxotrophic strains were generated to work with auxotrophic makers. 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]
In the peer-reviewed literature report, experimental results on function and interaction of yeast genes are extracted by high-quality manual curation and integrated within a well-developed database. The data are combined with quality high-throughput results and posted on Locus Summary pages which is a powerful query engine and rich genome browser.
The assay has a very good sensitivity (detection limit for 17β-estradiol in the YES assay about 5 x 10 −12 M or 1.4 ng/L), [1] [2] [4] and the microplate format requires only small amounts of sample. Analysis of native aqueous samples, concentrated environmental samples and chemicals or mixtures in solvents like ethanol or DMSO is possible.
Complementation tests can also be carried out with haploid eukaryotes such as fungi, with bacteria, and with viruses such as bacteriophage. [1] Research on the fungus Neurospora crassa led to the development of the one-gene-one-enzyme concept that provided the foundation for the subsequent development of molecular genetics.
The permissive temperature is the temperature at which a temperature-sensitive mutant gene product takes on a normal, functional phenotype. [2] When a temperature-sensitive mutant is grown in a permissive condition, the mutant gene product behaves normally (meaning that the phenotype is not observed), even if there is a mutant allele present.