PCV2衣壳蛋白羧基端基因在T4噬菌体表面的展示

用PCR扩增猪圆环病毒II型广东分离株的衣壳蛋白羧基端基因，将PCR产物连接到重组型质粒pR上，转化DH5α细胞并筛选阳性克隆；重组质粒经PCR鉴定并测序后，转化E2菌，将重组E2菌与缺陷型噬菌体T4-Z1同源重组后，得到重组噬菌体，SDS-PAGE和Westernbloting分析，表明衣壳蛋白基因在噬菌体表面正确展示，表达的融合蛋白的分子量约为25Ku。

Display of porcine circovirus type 2 cap gene on the T4 bacteriophage surface

Based on the gene sequence of cap of PCV2, a pair of primers was designed and, a 384 bp fraction of the carboxyl end of the capsid protein gene was obtained by PCR amplification, which was then cloned into plasmid pR to yield an integrative plasmid pR-cap. The integrative recombinant plasmid was used to transform E. coli E2 host bacteria, and then this E2 containing the recombinant plasmid was used for homologous recombination with lysozyme gene-deficient T4, thus yielding a recombinant bacteriophage T4-cap. When purified T4-CAP was tested with SDS-PAGE and Western blotting, a 25 Ku protein band was detected in polyacrylamide gel and nitrocelluLose membrane, which could combine specifically with PCV2 antiserum, attesting to the successful display of SOC fusion protein on the T4 bacteriophage surface.