摘要
本研究利用PCR方法扩增出Ⅰ群禽腺病毒FAVⅠhexon基因主要抗原区,连接于表达载体pGEX-4T-1中,构建成重组质粒pGEX-hexon,并转化大肠杆菌DH5α,用IPTG诱导表达,对表达的Hexon蛋白进行SDS-PAGE电泳和Western-blot分析。结果显示,试验成功扩增获得hexon基因主要抗原区序列,大小为1020bp。重组蛋白主要以包涵体形式存在,融合蛋白大小为63.1ku,与预测值相符。经Western-blot分析,表明重组蛋白与FAVⅠ的阳性血清发生特异性反应,具有良好的反应原性,为FAVⅠ诊断试剂盒的研制奠定基础。
The major antigenic region of fowl adenovirus group Ⅰ（FAVⅠ） hexon gene was amplified by PCR method,and was subcloned into prokaryotic vector pGEX-4T-1 to construct recombinant expression plasmid pGEX-hexon.The recombinant plasmid was transformed into Escherichia coli DH5α,and then induced and expressed by IPTG.The truncated Hexon recombinant protein was analyzed by SDS-PAGE and western-blot.The result showed that the major antigenic region of hexon gene with a size of 1 020 bp was successfully amplified.The truncated hexon recombinant protein was mainly expressed in the form of inclusion body and the molecular weight of the fusion protein was about 63.1 ku.The result of the western-blot showed that the truncated hexon recombinant protein could be specifically recognized by positive serum to FAVⅠ,and may be used as the coating antigen to develop a diagnosis kit against fowl adenovirus group Ⅰ.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2011年第5期550-555,共6页
Genomics and Applied Biology
基金
广西特聘专家专项经费和广西科技项目(桂科攻0815009-3-6,2010GXNSFA013090和桂兽研专项08-1)经费共同资助
