酵母菌Y12667 ERG11基因删除及验证研究  

Deletion and Verification of the Correct Recombination of the ERG11 Gene in Yeast Y12667

作　　者：陈双红[1,2] 盛春泉[2] 徐晓辉[2] 姜远英[2] 张万年[2] 何成[2] 

机构地区：[1]海军医学研究所,中国上海200433 [2]第二军医大学,中国上海200433

出　　处：《生命科学研究》2009年第2期116-121,共6页Life Science Research

基　　金：国家自然科学基金重点资助项目(30430750)

摘　　要：采用同源重组的方法删除酵母菌Y12667内源ERG11基因,并用PCR、Western、细胞计数和GC-MS的方法分别在基因水平、蛋白水平、细胞水平和功能代谢水平进行验证.结果表明,本研究成功删除了酵母菌Y12667内源ERG11基因,该基因删除菌能稳定表达外源性白念珠菌ERG11基因的同源蛋白.因此,该基因删除菌可作为下一步靶酶蛋白功能研究的基因工程表达操作菌.ERG11 gene of Saccharomyces cerevisiae was chosen as target gene to be disrupted. PCR-based gene deletion strategy was employed to knock out this gene. Targeting of the replacement cassettes at the genomic locus was verified by PCR analysis with the Zeocin gene as a probe. The inactive function of CYPS1 was observed through protein by Western blot and lanosterol composing by GC-GS. The result shows that, with the new deletion cassette, ERG11 gene has been disrupted successfully in Y12667.

关 键 词：酵母菌 白念珠菌 羊毛甾醇14α-去甲基化酶 基因删除 

分 类 号：Q784[生物学—分子生物学]