酵母醇脱氢酶辅酶结合结构域基因片段的克隆与表达  

Cloning of the Gene Fragment Encoding the CoenzymeBinding Domain of Yeast ADH I and Its Fusion Expression in E.coli

作　　者：廉德君[1] 许根俊[1] 

机构地区：[1]中国科学院上海生物化学研究所,中国科学院生物物理研究所生物大分子国家重点实验室

出　　处：《生物化学与生物物理学报》1996年第4期389-395,共7页

基　　金：国家自然科学基金

摘　　要：本文报道应用重组ＤＮＡ方法对酵母醇脱氢酶（ＹＡＤＨ）结构基因－功能片段的定向克隆与表达。应用聚合酶链式反应（ＰＣＲ）技术实现了对ＹＡＤＨ组成型同工酶ＡＤＨＩ结构基因中编码ＮＡＤ结合结构域的一段约４００ｂｐ的基因片段的定向克隆。序列分析表明得到的基因片段完全符合设计。表达在大肠杆菌中进行，所用的表达载体为大肠杆菌融合表达系统，融合蛋白的产率达细胞总蛋白的２０％。本工作为研究分离的结构域的结构与功能打下了基础。In this communication we report the cloning and the overexpression of the coenzyme-binding domain of yeast alcohol dehydrogenase.A gene fragment about 400 bp encoding the NAD binding domain of yeast S.cerevisiae alcohol dehydrogenase I was obtained by the PCR method.The domain was expressed as a fusion protein by a plasmid which was constructed by inserting the PCR-amplified gene fragment into the plasmid pWR590-1 which includes E.coli Lac promotor and a truncated β-galaetosidase gene.The fusion protein was characterized by SDSPAGE and Western blot analysis.The yield of the fusion protein was 20%of the total E.coli protein.

关 键 词：酵母 醇脱氢酶 结构域 基因克隆 高效表达 

分 类 号：Q949.326.1[生物学—植物学] Q554.9