Development of a highly efficient gene targeting system induced by transient repression of YKU80 expression in Candida glabrata.

ABSTRACT: In the pathogenic yeast Candida glabrata, gene targeting to generate knockouts and "knockins" is a potentially powerful method for the analysis of gene function. Its importance increased after the C. glabrata genome sequence project, but progress in the field is hampered by inefficient mechanisms for gene targeting. With the use of 40-bp homologous flanking DNA, no gene targeting was identified. To address this issue, YKU80 was disrupted, leading to an increase in targeting efficiency of 5.1% using 40-bp flanking homologous DNA. To harness the beneficial effects of YKU80 inactivation on gene targeting frequency without incurring any negative effects, such as synthetic sickness or lethality, we developed a new system whereby the expression of YKU80 was restored following a transient knockdo

SUBMITTER: Ueno K 

PROVIDER: S-EPMC1951112 | biostudies-literature | 2007 Jul

REPOSITORIES: biostudies-literature