GENERATION OF CONDITIONAL YEAST MUTANTS, METHODS AND REAGENTS RELATED THERETO

摘要

The invention features a method for systematically generating mutant strains of obligate haploid and obligate diploid yeast, having condition-sensitive lethal mutations of a gene which is otherwise essential to cell viability under all conditions of cell growth. Conditional mutants make possible the analysis of physiological changes caused by inactivation of a gene or a gene product and can be used to address the function of any gene. In an exemplary embodiment, the method of the present invention can be generally characterized as follows: (i) providing a Candida strain whichis heterozygous for a loss-of-function mutation of an allele of an essential gene; and (ii) introducing a conditionally sensitive homolog of the gene, e.g., with conditional-sensitive protein or conditional-sensitive transcriptional regulatory sequence, at the functional allele by integrative transformation.