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.
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)
