摘要
为原核表达基因C型鸭甲肝病毒(DHAV-C)VP1重组蛋白,本研究通过设计套式PCR引物,RT-PCR扩增DHAV-C VP1全基因,约为720 bp,将其亚克隆至pET-32a(+)载体中构建重组表达质粒pET-VP1。将其转化E.coli Rosetta(DE3)中,经IPTG诱导表达了约47 ku的重组蛋白。Western blot分析表明,该重组蛋白可以与兔抗DHAV-C阳性血清发生特异性反应,具有良好的反应原性。
To express duck hepatitis A virus genotype C （DHAV-C） VP1 gene, Nested-PCR primers were designed to amplify VP1 gene from DHAV-C strain by RT-PCR and cloned into pET-32a（＋） for expressing in E. coli recombinant VP1was highly expressed at 8 hour under 1 mM IPTG induction. SDS-PAGE showed the relative molecular weight of the VP1 recombinant protein was about 47 ku, and western blot analysis showed that the VP1 recombinant protein reacted with rabbit anti DHAV-C.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2012年第9期740-742,共3页
Chinese Journal of Preventive Veterinary Medicine
基金
西南民族大学中央高校专项资金项目(11ZYXS22)
关键词
基因C型鸭甲肝病毒
VP1基因
原核表达
duck hepatitis A virus genotype C
VP1 gene
prokaryotic expression
