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| 人/猴免疫缺陷病毒SHIV衣壳蛋白p27的表达纯化及活性鉴定 | |
| 引用本文： | 华秋东,姜成刚,王海,相文华,周建华,陈维多.人/猴免疫缺陷病毒SHIV衣壳蛋白p27的表达纯化及活性鉴定[J].东北农业大学学报,2008,39(11). |
| 作者姓名： | 华秋东 姜成刚 王海 相文华 周建华 陈维多 |
| 作者单位： | 1. 东北农业大学生命科学学院,哈尔滨,150030;中国农业科学院哈尔滨兽医研究所,哈尔滨,150001 2. 中国农业科学院哈尔滨兽医研究所,哈尔滨,150001 3. 东北农业大学生命科学学院,哈尔滨,150030 |
| 基金项目： | 黑龙江省科技攻关项目，哈尔滨市科技攻关项目 |
| 摘 要： | 利用PCR技术从质粒SHIV89.6P中扩增p27gag基因,并克隆入pMD18-T载体,测序鉴定;将p27gag基因插入表达载体pET32aT7启动子下游,构成重组质粒pET32a-p27并使其在大肠杆菌BL21中高效表达,最后利用固定化金属离子(Ni2+)配体亲和层析技术从表达蛋白中纯化目的蛋白,Western Blot检测活性。结果显示,融合蛋白p27经SDS-PAGE电泳观察可见45ku的明显条带,与预期分子质量大小一致且多以可溶形式表达,目的蛋白占菌体总蛋白的36.8%;经镍柱纯化,获得目的蛋白p27纯度可达98%;经Western Blot试验检测,证明此融合蛋白p27能与SHIV阳性血清发生特异性反应,具有较好的抗原活性。该研究为进一步开发早期诊断试剂盒和基因工程疫苗奠定了初步基础。 |
| 关 键 词： | SHIV p27 原核表达 大肠杆菌 |
Expression, purification and bioactivity test of SHIV capsid p27 |
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| HUA Qiudong,JIANG Chenggang,WANG Hai,XIANG Wenhua,ZHOU Jianhua,CHEN Weiduo.Expression, purification and bioactivity test of SHIV capsid p27[J].Journal of Northeast Agricultural University,2008,39(11). | |
| Authors: | HUA Qiudong JIANG Chenggang WANG Hai XIANG Wenhua ZHOU Jianhua CHEN Weiduo |
| Institution: | HUA Qiudong1,2,JIANG Chenggang2,WANG Hai1,XIANG Wenhua2,ZHOU Jianhua2,CHEN Weiduo1 (1. College of Life Sciences,Northeast Agricultural University,Harbin 150030,China,2. Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China) |
| Abstract: | According to the published SHIV sequences, p27 gag gene was amplified from the plasmid SHIV89.6P by PCR method, cloned into the pMD18-T vector and sequenced. Under the control of the bacterio- phage T7 promoter, the gag gene fragment of p27 was inserted into the pET32a vector in order to construct the pET32a -p27 recombined vector. After highly expresson in E.coli, the purified p27 protein was prepared by metal-ligand affinity chromatography(IMAC). The accuracy of the inserted gene and activity of the SHIV ... |
| Keywords: | SHIV p27 |
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