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猪瘟病毒NS3蛋白ATPase/RNA解旋酶功能区的表达与纯化
引用本文:郑杰,宁宜宝.猪瘟病毒NS3蛋白ATPase/RNA解旋酶功能区的表达与纯化[J].中国畜牧兽医,2007,34(10):43-46.
作者姓名:郑杰  宁宜宝
作者单位:1.中国农业大学动物医学院国家动物海绵状脑病实验室,北京 100094;2.中国兽医药品监察所,北京 100081
摘    要:采用RT-PCR 技术扩增出猪瘟病毒石门株和C株NS3基因的ATPase/RNA解旋酶功能区,将其克隆到表达载体pET-28a(+) 中,获得重组质粒pET-NS3SM和pET-NS3C;PCR、酶切鉴定和序列分析结果表明目的基因插入位置、方向和读码框完全正确;0.8 mmol/L IPTG诱导得到分子量为54kD 的目的蛋白,Western blotting结果表明,表达的目的蛋白与猪瘟高免血清没有出现肉眼可见的反应;通过ELISA试验结果表明,重组蛋白能被CSFV 阳性血清识别。

关 键 词:猪瘟病毒  NS3基因  ATPase/RNA解旋酶  克隆  表达    
文章编号:1671-7236(2007)10-0043-04
修稿时间:2007-04-18

Cloning Expression,Purification of Partial ATPase/RNA Helicase Functional Domain of NS3 Genes of Classical Swine Fever Virus
ZHENG Jie,NING Yi-bao.Cloning Expression,Purification of Partial ATPase/RNA Helicase Functional Domain of NS3 Genes of Classical Swine Fever Virus[J].China Animal Husbandry & Veterinary Medicine,2007,34(10):43-46.
Authors:ZHENG Jie  NING Yi-bao
Affiliation:1.National Animal Transmissible Spongiform Encephalopathies Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100094,China;2.National Classical Swine Fever Reference Laboratory,China Institute of Veterinary Drug Control, Beijing 100081,China
Abstract:Partial ATPase/RNA helicase functional domain of NS3 genes of CSFV SHIMEN rain and C rain were amplified by RT-PCR;The recombinant plasmids pET-NS3 SMand p ET-NS3C were obtained after being cloned them into expression vector pET-28a( );And then the 54 kD target proteins were produced by inducing with 0.8 mmol/L IPTG;Western blotting showed that the expressed proteins couldn't be recognized by superserum against csfv.And ELISA showed that the expressed proteins could be recognized by superserum against csfv.
Keywords:classical swine fever virus(CSFV)  NS3 gene  ATPase/RNA helicase  cloning  expression
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