作　　者： ; ; ; ; (崔东）;

机构地区： 华南理工大学生物科学与工程学院生物工程系

出　　处： 《华南理工大学学报（自然科学版）》 2004年第6期89-92,共4页

摘　　要： 将携有mTn-3xHA/LacZ转座子的酿酒酵母文库质粒pHSS6转化大肠杆菌DH5α,挑取单菌落并提取其质粒.将纯化质粒分别用Not I酶切后,转化尿嘧啶缺陷型(ura-)酿酒酵母菌株INVSc1,使其同源重组到酿酒酵母基因组中,于SC/ura-平板上筛选.取长有单菌落酵母的平板进一步筛选重组后上游有强启动子酿酒酵母文库质粒,用近300个质粒转化酵母菌株INVSc1,最终得到2株前端有强启动子的酿酒酵母质粒,这2个质粒可作为酿酒酵母同源重组载体广泛应用. Firstly, Saccharomyces cerevisiae library plasmids pHSS6-mTn-3xHA/LacZ were transfected into E. coli DH5α, and isolated plasmids were selected from single colonies. Next, the purified plasmids were individually digested with Not I and were used to transfect Saccharomyces cerevisiae INVScl with uracil auxotrophy (ura-) in order to recombine the plasmids with Saccharomyces cerevisiae genomes homologously. Then, SC/ ura- plates were used to screen positive recombinant clones that were further screened from the plates with single colonies, and strong promoters upstream the LacZ gene were found. Finally, nearly 300 plasmids were transfected into the INVScl strain of yeast, and two strong upstream promoter-containing plasmids were isolated, which can be widely used as the homologous recombinant vectors of Saccharomyces cerevisiae.

关 键 词： 酿酒酵母 同源重组 转座子 化学转化 同源重组载体

领　　域： [生物学]