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Cloning and Expression of Tibetan Sheep-origin Echinococcus granulosus Antigen B Gene and Protein Identification#br#  Using Immunological Method

DUO Hong1, LI Wei1, FU Yong1, PENG Mao1, GUO Zhi-hong1，SHEN Xiu-ying1, #br# NIU Jian-zhang1, NI Ma2, TONG Yan-jun3, HE Sheng-de2, E Ri-jie3,#br# CHUI Wen-ting4, HU Bao-ping4   

  1. 1 Qinghai Academy of Animal Science and Veterinary Science, Qinghai 810016, China；2 Haiyan County Veterinary Station of Qinghai Province, Haiyan 812200, China；3 Xinghai County Veterinary Station of Qinghai Province, Xinghai 813300, China；4 Qinghai University, Qinghai 810016, China
  • Online:2016-02-28 Published:2016-03-11

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Abstract:

Objective To clone and express the Tibetan Sheep-origin Echinococcus granulosus Antigen B8/2 Gene, and immunologically identify the encoded protein. Methods The cDNA of EgAgB8/2 gene was amplified by RT-PCR. The prokaryotic expression vector pET-EgAgB8/2 was constructed and transformed into E. coli BL21（DE3） for expression. Proteins were extracted, separated in SDS-PAGE and identified by Western blotting. Results The cloned EgAgB8/2 gene was 335 bp in length, and had a 98%-100% sequence homology with the reported cDNA sequence of EgAgB8/2, indicating the successful construction of the pET-EgAgB8/2 vector. SDS-PAGE revealed large amount of proteins in supernatant. Western blotting further confirmed the expression of the target protein. Conclusion  The EgAgB8/2 gene of Tibetan Sheep-origin in Qinghai is successfully cloned, and the constructed pET-EgAgB8/2 vector can be used to express the target protein.

Key words: Echinococcus granulosus, AgB8/2 gene, Gene cloning, Prokaryotic expression