猪细小病毒(PPV)SD1株NS1基因的克隆与原核表达

为研究猪细小病毒（Porcine Parvovirus，PPV）NS1基因，建立快速、准确诊断猪细小病毒的方法，减少猪细小病毒造成的损失，笔者克隆了含有NS1基因的片段，并且与已登录猪细小病毒的相应序列进行比较，成功构建了原核表达重组质粒PET 30a／NS1，并在E.coli中进行了诱导表达。

Cloning and Prokaryotic Expression of NS1 Gene of Porcine Parvovirus(PPV)SD1 Strain

[Objective]The research aimed to provide the theoretical basis for establishing a rapid diagnosis method for porcine parvovirus(PPV).[Method]One pair of primers were designed according to PPV genome sequences on GenBank website and the sequences of prokaryotic expression vector pET30a( )with multiple cloning sites. The whole sequence of NS1 gene in PPV SD1 strain Was amplified by using PCR technology and the positive recombinant plasmid was analyzed by sequencing and homology comparison.The prokaryotic expression recombinant plasmid PET30a/NS1 was constrocted to make its induction expression in Escherichia coli.[Result]The target fragment with the length of 2 208 bp was obtained from PCR amplification.The nucleotide homologies between the cloned NS1 gene and the reported relevant PPV genes were from 97.3% to 99.4%.which indicated that NS1 gene had high conservation.But it had a 12-basepair successive deletion near the hydroxyl end.The cloned PPV NS1 gene was successfully expressed in prokaryotic cell,and its expression products existed mostly in inclusion bodies. [Conclusion]The results of SDS-PAGE detection showed that the molecular weight of PPV NS1 protein was 86 KD.