番鸭细小病毒VP3基因的原核表达及鉴定

目的]使番鸭细小病毒(DPV)VP3在原核表达系统中正确表达.方法]根据番鸭细小病毒VP3基因序列,设计一对特异性引物,利用PCR技术扩增出VP3基因;将其克隆至原核表达载体p ET-32a,获得重组表达载体p ET-32a-VP3.将重组质粒转化感受态细胞BL21(DE3),经IPTG诱导后,SDS-PAGE检测重组蛋白.结果]成功表达出与预期大小相符的重组蛋白,约89 k Da;且当IPTG浓度为1.2 mmol/L,诱导时间为6 h时蛋白表达量最大.结论]该研究通过原核表达系统成功表达了DPV、VP3蛋白,并摸索了蛋白最佳表达条件.

Prokaryotic Expression and Evaluation of VP3 Gene of Muscovy Duck Parvovirus

Objective]To prepare the prokaryotic expression of VP3gene of Muscovy duck parvovirus. Method]One pair of specific primers was designed to amplify VP3 gene by PCR. The amplified fragments were cloned into prokaryotic expression vector pET-32a,and the recombinant plas-mids were transformed into E. coli BL21(DE3)cells. The protein was expressed following IPTG induction,and detected by the SDS-PAGE. Re-sult]The recombinant protein matched with the expected result was successfully expresssed,the size of the recombinant protein was about 89 kDa. When the concentration of IPTG was 1. 2 mmol/L,the time was 6 h,the expression of protein was the largest. Conclusion]The expression of VP3 protein of DPV was successfully expressed through the prokaryotic expression system,and the best expression conditions were explored.