Welcome to Chinese Agricultural Science Bulletin,

Chinese Agricultural Science Bulletin ›› 2012, Vol. 28 ›› Issue (20): 98-101.doi: 10.11924/j.issn.1000-6850.2012-1656

Special Issue: 生物技术 畜牧兽医

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Construction of Prokaryotic Expression Vector of Cap and Rep Protein of Porcine Circovirus Type Ⅱ and Expression in E.coli BL21

  

  • Received:2012-04-28 Revised:2012-05-07 Online:2012-07-15 Published:2012-07-15

Abstract:

In order to develop effective detection methods and prepare monoclonal antibody of porcine circovirus type Ⅱ , Cap and Rep genes of porcine circovirus type Ⅱ were amplified and cloned into pMD18-Tsimple vector. And the genes were sequenced. Then Cap and Rep genes were sub-cloned into pACYCDuet-1 vector to generate three prokaryotic expression plasmids named pACYCDuet-1-Cap, pACYCDuet-1-Rep, and pACYCDuet-1-Rep -Cap. The plasmids were transformed into E.coli BL21 (DE3) and expressed. The results of Western-blotting showed that the two proteins were expressed successfully in E.coli BL21 (DE3). Furthermore, plasmid for coexpression of two proteins expressed better than the other two plasmids.