PCV2ORF1、ORF2截短基因原核表达载体的构建及表达  

Cloning and Prokaryotic Expression of the ORF1 and the ORF2 Truncated Genes of Porcine Circovirus Type 2

作　　者：时建立[1,2] 李俊[1,2] 丁鹏[3,2] 吴家强[1,2] 孙文博[1,2] 丛晓燕[1,2] 周顺[3] 

机构地区：[1]山东省农科院畜牧兽医研究所,山东济南250100 [2]山东省畜禽疫病防治与繁育重点实验室,山东济南250100 [3]青岛农业大学,山东青岛266109

出　　处：《家畜生态学报》2010年第3期24-26,共3页Journal of Domestic Animal Ecology

基　　金：山东省农业科学院青年基金项目(2006YQN033);山东省农业科学院创新基金重点项目课题(2007YCX017-4);山东省自然基金项目(Y2006D23);山东省攻关项目(2008GG10009034)

摘　　要：采用PCR方法,从PCV2双拷贝DNA感染性克隆中扩增出PCV2 ORF1和ORF2截短基因,与pQE30原核表达载体分别经KpnI和HandIII双酶切,连接,经鉴定后证明成功构建pQE30-ORF1、pQE30-ORF2原核表达载体,经IPTG诱导,SDS-PAGE和West-ern-blot检测到目的蛋白表达,为研究蛋白功能和单克隆抗体的研制打下了基础。PCV2ORF1 and ORF2truncated genes were amplified by PCR from the double-copies of infectious DNA clone and inserted into the prokaryotic expression vector pQE30 used double restriction-enzyme Kpn I and Hand III digestion respectively.The recombinants were constructed successfully used PCR、double restriction-enzyme digestion and sequenced.Purpose proteins were detected expression by SDS-PAGE and Western-blot after induced by IPTG.The results suggest an foundation for protein function and Mcab research.

关 键 词：PCV2 ORF1 ORF2 截短 表达 

分 类 号：S811.5[农业科学—畜牧学]