| 脱水蛋白DHN1表达载体pBV221-dhn1的构建及基因表达 |
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| 引用本文： | 张永炜,邱全胜.脱水蛋白DHN1表达载体pBV221-dhn1的构建及基因表达[J].北京师范大学学报(自然科学版),2001,37(2):250-254. |
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| 作者姓名： | 张永炜 邱全胜 |
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| 作者单位： | 北京师范大学生命科学学院, |
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| 基金项目： | 国家重点基础研究发展计划(973计划);G1999011705; |
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| 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.
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| 关 键 词： | 脱水蛋白 表达载体 PCR 基因表达 热溶性分析 |
| 修稿时间： | 2000年11月1日 |
THE CONSTRUCTION OF DEHYDRIN EXPRESSION VECTOR pBV221-dhn1 AND ITS GENE EXPRESSION |
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| ZHANG Yongwei,Qiu Quansheng.THE CONSTRUCTION OF DEHYDRIN EXPRESSION VECTOR pBV221-dhn1 AND ITS GENE EXPRESSION[J].Journal of Beijing Normal University(Natural Science),2001,37(2):250-254. |
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| Authors: | ZHANG Yongwei Qiu Quansheng |
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| Abstract: | 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 516bp fragment with the same size and sequence of predicted dhn1 is obtained. Expression study shows that a specific protein of 22×103 with the characteristics of heat resistance is produced after gene transformation. Taken together, a highly expressed dehydrin expression vector pBV221-dhn1 is obtained. |
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| Keywords: | dehydrin expression vector PCR gene expression analysis of high-temperature solubility |
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