›› 2013, Vol. 31 ›› Issue (4): 9-290-292.

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Cloning and Prokaryotic Expression of Casein Kinase Ⅱ Subunit Beta Gene Fragment of Dirofilaria immitis

GAO Jun-shan,WU Wei, HOU Hong-lie, GONG Peng-tao, LI Jian-hua, LI He, ZHANG Guo-cai, ZHANG Xi-cheng, YANG Ju*   

  1. College of Veterinary Medicine,Jilin University,Changchun 130062,China
  • Online:2013-08-30 Published:2013-12-27

Abstract:  Objective  To clone and express the partial fragment of Csnk2b gene of Dirofilaria immitis in prokaryotic cells, and analyze the immunoreactivity.  Methods  The partial fragment of Csnk2b gene was amplified by PCR with a pair of specific primers. The PCR product was cloned into pMD18-T, and then sub-cloned to pGEX-4T-1 expression vector. The constructed plasmid pGEX-4T-1-Csnk2b was transformed into E. coli Rosetta(DE3) and followed by expression of the protein induced by IPTG. The recombinant protein was analyzed by SDS-PAGE and identified by Western blotting.  Results  The PCR product was about 700 bp. Enzyme digestion and DNA sequencing confirmed that the recombinant plasmid pGEX-4T-1-Csnk2b was constructed. SDS-PAGE results showed that the relative molecular weight (Mr) of the fusion protein(GST-Csnk2b) was about 45 000. GST-Csnk2b reacted positively with mouse anti-D. immitis serum.  Conclusion  The partial Csnk2b gene has been expressed in prokaryotic expression system and shows immunoreactivity.

Key words:  , Dirofilaria immitis;Csnk2b;Prokaryotic expression