earticle

영어

Cells can repair DNA double-strand breaks by both homologous and nonhomologous mechanisms. Nonhomologous end joining (NHEJ), the direct rejoining of DNA double-strand breaks, is regulated by KU70/KU80 proteins and its related components. While Kluyveromyces marxianus is a promising yeast strain for biotechnological applications, genetic engineering of this strain is still challenging. Unusually, NHEJ frequency of K.marxianus is higher then Saccharomyces cerevisiae and other yeast strains. This high frequency of NHEJ could be advantageous for multiple gene integration. But, the high NHEJ interfere with homolgous (targeted) integration preventing targeted gene knock-out. In order to engineer the NHEJ and HR of K.marxianus regulatable, we substituted the promoter of ku70 with those of Gal which can make inducible expression of ku70. This strain enabled controllable gene introduction by switching NHEJ (one-step multiple integration) and homologous recombination (targeted gene insertion/knock-out).