猪瘟病毒NS3蛋白ATPase/RNA解旋酶功能区的表达与纯化  

Cloning Expression,Purification of Partial ATPase/RNA Helicase Functional Domain of NS3 Genes of Classical Swine Fever Virus

作　　者：郑杰[1] 宁宜宝[2] 

机构地区：[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

出　　处：《中国畜牧兽医》2007年第10期43-46,共4页China Animal Husbandry & Veterinary Medicine

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。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 esfv. And ELISA showed that the expressed proteins could be recognized by superserum against esfv.

关 键 词：猪瘟病毒 NS3基因 ATPase/RNA解旋酶 克隆 表达 

分 类 号：S852.651[农业科学—基础兽医学]