猪圆环病毒3型陕西株Cap基因的克隆、序列分析及原核表达

为了克隆猪圆环病毒3型Cap基因并进行序列分析和原核表达,参考GenBank上已发表的PCV3(KY418606.1)Cap基因序列,合成1对上、下游引物,PCR扩增Cap基因,进行序列分析和原核表达。结果显示,成功克隆到642 bp的Cap基因,核苷酸序列分析显示,PCV3 Cap基因与国内分离株核苷酸同源性最高为95.97%,氨基酸分析显示Cap蛋白不含信号肽和跨膜区。经IPTG诱导后获得了分子质量约35 ku的Cap蛋白,该蛋白能与PCV3阳性血清结合,具备良好的抗原性。成功克隆并原核表达PCV3陕西株Cap基因,为进一步建立PCV3的快速检测方法和研制PCV3亚单位疫苗奠定了基础。

Cloning,Sequence Analysis and Prokaryotic Expression of Cap Gene of Porcin Circovirus Type 3 Shaanxi Strain

The cloning,sequence analysis and prokaryotic expression of the porcine circovirus type 3 Shaanxi isolate Cap gene were performed.A pair of upstream and downstream primers were synthesized by referring to the sequence of PCV3 Cap gene registered in GenBank.The Cap gene was amplified by PCR and analyzed by sequence analysis and prokaryotic expression.642 bp Cap gene of PCV3 was successfully cloned.The nucleotide sequence analysis showed that the Cap gene nucleotide homology of PCV3 Shaanxi strain and domestic isolate was 95.97%.Amino acid analysis indicated that cap protein contained no signal peptide and transmembrane region.After induction by IPTG,the Cap protein with a molecular weight of 35 ku was successfully expressed.The Cap protein could react with PCV3 positive serum and had good antigenicity.In this study,Cap gene of PCV3 Shaanxi strain of was successfully cloned and expressed in prokaryotes,laying a foundation for the further establishment of a rapid diagnostic method for PCV3 and the development of PCV3 subunit vaccine.