鹅副粘病毒M基因主要抗原位点片段的原核表达及鉴定

Prokaryotic expression of major antigen site in M gene from goose paramyxovirus and detection

根据GenBank中发表的GPMV SF02株M基因核苷酸序列设计合成一对引物,通过RT-PCR扩增JS/1/97/G0株M基因主要抗原位点片段M1,将其克隆入pMD18-T载体,序列测定和分析表明,所扩增的M1基因核苷酸长为444 bp,共编码148个氨基酸。将M1同载体PGEX-6P-1连接后,转化入E.coliBL21(DE3)PlysS,经IPTG诱导表达出目标蛋白,与预计相符。对表达的M1蛋白进行Western blot鉴定,表明所表达的M1蛋白可以与GPMVSF02株阳性血清发生特异性反应。

In this study, according to M gene sequence of GPMV SF02 isolate strain published in GenBank, two pairs of primers were desighed and M1 gene was amplified by RT- PCR. The specific DNA products were cloned into pMD18-T vector. The sequence analysis showed that M1 gene was 444 bp in length, encoding 148 amino acid residues; the major antigen site of M gene was inserted into expression plasmid pGEX-6P-1. The recombinant plasmid pGEX-6P-M1 was transformed into E.coli BL21（DE3）PlysS and induced with IPTG. A fusion protein as we expected had been found. The expressed products were tested by western blot. The results showed that GST-M1 fusion protein had a positive reaction.