猪生殖与呼吸综合征病毒N基因的扩增与克隆

应用RT PCR方法扩增出猪生殖与呼吸综合征病毒 (PRRSV)的核衣壳蛋白基因 (N基因 ) ,并将其克隆到 pET 32a载体 ,构建了高效原核表达载体pETN。将 pETN重组质粒转化BL2 1(DE3)宿主菌后 ,对插入片段进行了序列测定及同源性分析。结果表明 ,插入片段序列与PRRSV美洲型ATCCVR2 332株的ORF7核苷酸序列同源性达 99.9% ,与分离毒株的同源性达 99.0 %。

Amplification and cloning of nucleocapsid protein gene of PRRSV

The nucleocapsid protein gene (N gene) of porcine reproductive and respiratory syndrome virus (PRRSV) was amplified by RT-PCR. After the gene has been cloned into pET-32a prokaryotic vector, a recombinant plasmid pETN was constructed. The recombinant plasmid was transformed into host cell BL21 (DE3), then sequenced and compared with other PRRSV isolates. The result showed that the nucleotide homology between the cloned N gene and American strain PRRSV ATCC VR2332 N gene was 99.9% and the nucleotide homology between the cloned N gene and the PRRSV isolate N gene was (99.0%.)