鹅副黏病毒L基因片段的克隆及其主要抗原表位区的原核表达

Construction of pGEX-6p-1-L Prokaryotic expression vector and expression of the major sequence of L gene from goose paramyxovirus

利用PCR扩增鹅副黏病毒编码L蛋白的主要抗原表位区基因片段,并将该片段克隆至原核表达载体pGEX-6P-1,通过诱导表达条件的摸索,实现了L蛋白主要抗原表位区在大肠杆菌中的高效中的表达,SDS-PAGE检测表达的融合蛋白主要以可溶形式存在,Western blot分析重组蛋白具有良好的抗原性,并利用亲和层析得到融合蛋白。

To establish a prokaryotic expression system expressing major epitope domain of canine parvovirus L protein to serve the diagnosis of GPMV, The fragment of GPMV L gene was cloned by RT-PCR. Then recombi- nant E. coli strain BL-21 was induced to express the fusion protein by IPTG. The expressed products were detected by SDS-PAGE and western blot. The results showed that the fusion protein was efficiently expressed and the size of recombinant protein is about 47 kD, complying with expectations and has very good antigenicity.