玉米叶绿体偶联因子ε亚基基因在E．coli中的表达  

Expression of the atpE Cene Coding for the ε Subunit of Maize Ckloroplast Coupling Factor

作　　者：史瑾[1] 魏家绵[1] 沈允钢[1] 

机构地区：[1]中国科学院上海植物生理研究所

出　　处：《生物化学与生物物理学报》1996年第1期1-7,共7页

基　　金：国家自然科学基金;植物分子遗传国家重点实验室基金

摘　　要：玉米叶绿体偶联因子ε亚基基因（ａｔｐＥ）被克隆到载体ｐＪＬＡ５０５和ｐＷＡ的多聚接头处，形成重组质粒ｐＪＬＡ５０５－ａｔｐＥ和ｐＷＡ－ａｔｐＥ，转化Ｅ．ｃｏｌｉ，并进行温敏诱导表达研究。在大肠杆菌中温敏诱导的基因表达产物经ＳＤＳ－ＰＡＧＥ测定，表达量达全菌蛋白质的３０％以上．Ｗｅｓｔｅｒｎ－ｂｌｏｔ分析结果表明温敏诱导的表达产物可特异性地与ε亚基抗体反应，在免疫双扩散中也观察到沉淀线。大肠杆菌表达的玉米叶绿体ａｔｐＥ基因产物以包含体形式存在于细菌中。经对包含体处理后；获得的粗产物可达８０％以上纯度，并具有天然ε亚基蛋白质的生物功能．The entire atpE gene of the maize ehloroplast coupling factor was inserted into the polylinker region of vectors PJLA505 and pEA to form recombinant plaamids pJLA505-atpE and pWA-atpE respectively.These expression plasmids were transformed into E.coli NM522 which induced at 42℃. By the analysis of SDS-PAGE, the empressed product of interest was observed to account for more than 30% of total E. coli cell proteins. The identification of the expressed.demonstrated that its immunological specificity was well retained. The antiserum cross-reacted with tie expressed ε protein and Cf1-ε protein of spinach and produced precipitin lines on Ouchterlony immunodiffusion test. The expressed product aggregated insolubly as the inclusion body and was purified to over 80%purity. The purified product had the εame function as that of the native εsubunit.

关 键 词：玉米 叶绿体偶联因子 Ε亚基 基因表达 

分 类 号：S513.01[农业科学—作物学]