摘要
利用RT-PCR技术扩增出不含信号肽的猪瘟病毒石门株的E2基因,将其克隆到PGEX-T-Easy 载体上,用BamH Ⅰ和HindⅢ进行双酶切,回收目的基因。将目的基因克隆到表达载体质粒pPROEX-HTb中,获得 重组质粒PPRO,EXE2,用pPROEXE2转化大肠杆菌,诱导含重组质粒PPROEXE2的大肠杆菌BL21表达E2基因蛋 白,研究E2蛋白与猪瘟阳性血清反应的特异性。结果表明,受体菌诱导后能表达E2基因蛋白,所表达的E2蛋白能 与猪瘟阳性血清发生特异性很强的反应。
The E2 gene of CSFV Shimen strain was amplified by reverse transcription polymerase chain reaction （RT-PCR）. The amplified fragments were cloned into PGEX-T-Easy vector. We had digested the interested gene by the property without both of the two cleavage sites of PGEX-T-Easy vector and obtained the recombinant plasmids pPRoEXE2 by cloning the interested gene into the expressed vector plasmids pPkoEX-HTb. It was identified that the position of the E2 gene insert,the size and the reading frame were all right by PCR,restriction digestion and the sequence analysis. SDS-PAGE indicated that the recipi ent germs induced by the recombinant plasmids could express E2 gene of CSFV. Western blot indicated that the positive serum of CSFV could recoznize the exoressed antigen protein.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2005年第12期13-16,共4页
Journal of Northwest A&F University(Natural Science Edition)
基金
陕西省科技攻关项目(2003K02-G11)
关键词
猪瘟病毒石门株
E2基因
克隆
表达
classical swine fever virus shimen strain
E2 gene
cloning
expression
