摘要
本研究基于犬细小病毒(canine parvovirus,CPV)VP2基因序列设计引物,利用PCR技术扩增该基因。扩增的VP2基因连入表达载体pET-32a(+),经PCR及双酶切鉴定基因连入正确。重组表达载体转入大肠杆菌后,经IPTG诱导,SDS-PAGE及Western blotting分析表明有特异蛋白的表达,分子质量大小为87 ku。将表达的蛋白包被酶标板后用抗犬细小病毒单抗与其反应,表明表达的VP2蛋白可以与相应抗体特异结合。
One specific pairs of primers,which were designed by canine parvovirus（CPV） VP2 gene sequence,were used to clone the VP2 gene isolated from Beijing through PCR.VP2 gene was subcloned from pEASY-T-VP2 into prokaryotic expression vector pET-32a.Sequence analysis confirmed that the VP2 gene was inserted correctly.SDS-PAGE and Western-blotting analysis revealed that the VP2 protein was expressed at high level in Escherichia coli.Through ELISA assay,the expressed VP2 protein has fine immunogenicity.
出处
《中国畜牧兽医》
CAS
北大核心
2010年第4期185-188,共4页
China Animal Husbandry & Veterinary Medicine
基金
中央级公益性科研院所基本科研业务费专项资金(2009QN-3)
关键词
犬细小病毒
表达
抗原性
canine parvovirus
expression
immunogenicity
