摘要
根据发表的犬瘟热病毒(CDV)参考株Ondetstepoort的序列设计一对引物,以CDV感染的Vero细胞总RNA为模板,经RT-PCR扩增得到549bp DNA片段,将该片段以正确的阅读框架定向克隆于pGEX-4T-1中,然后将重组质粒转化宿主菌RosettaTM,在37℃、1.0mmol/L IPTG诱导下外源基因获得良好表达。经SDS-PAGE鉴定,表达的融合蛋白约46ku,与预期值一致。Western blot试验显示,CDV免疫的小鼠血清可特异的识别该重组蛋白,表明该重组蛋白具有一定的免疫原性。
A 549 bp Haemagglutinin （H） gene fragment of CDV was amplified by polymerase chain reaction （PCR） using a pair of primers designed based on the H gene sequence of the Ondetstepoort strain, The PCR product was cloned into the expression plasmid vector pGEX-4T-1, then transformed into the RosettaTM and induced by 1.0 mmol/L IPTG at 37 ℃. The expression products were identified by SDS-PAGE and Western blot. The results showed that the H gene fragment was expressed as a 46ku protein which maintained some immunogenicity of CDV.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2008年第8期601-604,611,共5页
Chinese Journal of Preventive Veterinary Medicine
