根癌农杆菌介导酿酒酵母的遗传转化

Research of Agrobacterium tumefaciens-Mediated Transformation of Saccharamyces cerevisiae

根癌农杆菌(Agrobacterium tumefaciens)可以将它的Ti质粒的一段T-DNA序列转化到广泛的宿主细胞并能整合到宿主染色体中。作者利用根癌农杆菌转化系统成功地实现了产甘油假丝酵母(Candida glycerinogenes)乳清苷酸脱羧酶基因(Ura3)对酿酒酵母(Saccharamyces cere-visiae)W303-1A的Ura3缺陷遗传互补转化,转化率约为3.2个转化子/105个酵母细胞。通过转化子PCR和表型验证说明了T-DNA已经整合进入酵母染色体,并能够稳定地进行遗传表达。

Agrobacterium tumefaciens could transfer part of its Ti plamid and T-DNA to a wide variety of host cells where they could be integrated into the nuclear genome. Using this transformation system, the transformants were selected from SM （selected medium） plate with an efficiency of 3. 2 transformants per 10^5 yeast cells. The PCR and the phenotype results demonstrated the Ura3 gene has been integrated into chromosome of S. cerevisiae W303-1A successfully.