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Cloning and Prokaryotic Expression of Aeromonas veronii Flagellin flaA

ZHAI Xin-xin1,WEN Zhen-cai1,SHEN Xue-fei1,SUN Zhen1,JIA Sheng-mei1,ZHANG Jun-hui1,WANG Wen-dong2,YANG Zhen-guo1,LU Qiang1   

  1. 1.Institute of ZoonosesJilin UniversityChangchun 130062China2.College of Animal Science and Veterinary MedicineJilin UniversityChangchun 130062China

  • Revised:2014-04-10 Online:2014-07-20 Published:2014-08-21

Abstract: This study was aimed to express Aeromonas veronii flagellin flaA in E.coli BL21(DE3) and preliminarily identify antigenicity of recombinant protein through Western blotting.A pair of primers was designed according to the published sequences in GenBank.Complete ORF sequence of flaA gene was obtained by PCR amplification and cloned into pET-30a vector,and then transformed into E.coli BL21(DE3) to induce,express and purify.SDS-PAGE results showed that the target protein was successfully expressed in E.coli BL21(DE3) and the product was consistent with the theoretical value and seen clearly at approximately 38 ku.Western blotting result showed that the expression of recombinant proteins could produce immune reaction with prepared antibody and His-tag antibody.Therefore,Aeromonas veronii flagellin flaA was successfully expressed, which laid the basis for further study the role of biological function and immune adjuvant of flagellin.

Key words:

Aeromonas veroniimso-bidi-font-family: 宋体">;flagellincloningexpression