脱水蛋白DHN1表达载体pBV221-dhn1的构建及基因表达  

THE CONSTRUCTION OF DEHYDRIN EXPRESSION VECTOR pBV221-dhn1 AND ITS GENE EXPRESSION

作　　者：张永炜[1] 邱全胜[1] 

机构地区：[1]北京师范大学生命科学学院,北京100875

出　　处：《北京师范大学学报（自然科学版）》2001年第2期250-254,共5页Journal of Beijing Normal University(Natural Science)

基　　金：国家重点基础研究发展规划资助!项目 (G19990 1170 5 );国家自然科学基金!资助项目 (39770 0 77) ;教育部重点科技资助!项目 (99133)

摘　　要：以克隆载体 pTZ 19R dhn1(ZM )为基础 ,构建脱水蛋白DHN1表达载体 pBV2 2 1 dhn1.采用PCR技术从克隆载体 pTZ 19R dhn1(ZM )上扩增dhn1片段 ,并引入NcoⅠ /BamHⅠ酶切位点 ,然后与 pBV2 2 1原核表达载体连接 ,得到 pBV2 2 1 dhn1表达载体 .阳性克隆经PCR和NcoⅠ /BamHⅠ酶切检测都得到 516bp的dhn1片段 ,且序列正确 .表达载体pBV2 2 1 dhn1转化宿主菌后能够表达产生相对分子质量为 2 2× 10 3的DHN1,该蛋白具有高温可溶性 .以上结果表明 ,本实验得到了高效表达的脱水蛋白DHN1表达载体 pBV2 2 1 dhn1.pBV221-dhn1, an expression vector for dehydrin DHN1, is constructed from its cloning vector pTZ-19R-dhn1 (ZM) and its gene expression is studied .The dhn1 fragment with NocⅠ/BamHⅠrestriction sites is generated by PCR through cloning vector pTZ-19R-dhn1 (ZM), and cloned into the expression vector pBV221 with T4 ligase to produce the expression vector pBV221-dhn1. After transformation, positive clones are analyzed by PCR amplification and restriction enzyme analysis, and a 516 bp fragment with the same size and sequence of predicted dhn1 is obtained. Expression study shows that a specific protein of 22×10 3 with the characteristics of heat resistance is produced after gene transformation. Taken together, a highly expressed dehydrin expression vector pBV221-dhn1 is obtained.

关 键 词：脱水蛋白 表达载体 PCR 基因表达 热溶性分析 

分 类 号：Q753[生物学—分子生物学]