猪圆环病毒2型衣壳蛋白的表达

Expression of Capsid Protein in Porcine Circovirus Type 2

[目的]检测猪圆环病毒2型(PCV2)核衣壳蛋白(Cap)的表达,为进一步研制PCV2单克隆抗体检测抗原提供参考依据。[方法]根据已经发表的PCV2的衣壳蛋白基因(Cap)设计1对引物,利用PCR方法从已知PCV2病毒中扩增到1条DNA片段。将PCR产物按预定的阅读框架插入表达载体质粒pET28a(+)中,获得重组质粒pCAP-PCV2,并转化到大肠杆菌BL21(DE3)中。[结果]序列测定表明,所克隆的序列大小为579 bp,与DQ104422的Cap基因序列一致;通过对菌体裂解物的SDS-PAGE分析证明,携带重组质粒pET-PCV2-Cap的大肠杆菌可以表达可溶性的衣壳蛋白。[结论]PCV2衣壳蛋白能通过表达载体质粒pET28a(+)进行可溶性表达。

[Objective] To detect the expression of nucleocapsid protein（Cap） Porcine circovirus type 2（PCV2）,and provide a reference for the further development of antigen monoclonal antibodies in PCV2.[Method] One DNA fragment of Cap gene in PCV2 was amplified repectively by PCR using specific primers.After the DNA fragments was inserted reading frame pET28a（＋） plasmid expressing carrier,Recombinant plasmid pCAP-PCV2 was transformed into Escherichia coli BL21（DE3）.[Result] The sequence size was 579 bp and shared 100% identity with that of DQ104422.SDS-PAGE and Western blot analysis showed that the soluble cap protein was expressed.[Conclusion] PCV2 cap proteins can be expressed by the pET28a（＋） carrier.