›› 2010, Vol. 28 ›› Issue (2): 9-128.

• 论著 • Previous Articles     Next Articles

Prokaryotic Expression and Immunologic Identification of P-29 Gene from Echinococcus granulosus Hydatid Cyst

LI Jie1，WANG Zhi-gang1 *，HAO Xi-yan1，CHEN Xian-wei1，WANG Hong-xia1，LI Shu-yu1，YANG Jiao-fu1，YANG Jun2   

  1. 1 College of Life Science，Inner Mongolia University，Key Laboratory of Mammal Reproductive Biology and Biotechnology，Ministry of Education，Hohhot 010021，China；2 Hohhot Institute of Environmental Sciences，Hohhot 010030，China
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-04-30 Published:2010-04-30
  • Contact: WANG Zhi-gang

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Abstract: Objective To clone and express P-29 gene of Echinococcus granulosus, and analyze its immunore-activity. Methods Total RNA was extracted from the hydatid cyst of E. granulosus and its P-29 gene was amplified by RT-PCR. The PCR product was cloned into pET44a（+） vector. The recombinant plasmid was transformed into E. coli BL21 （DE3） and followed by expression of the protein induced by IPTG. The protein was purified, and tested by SDS-PAGE. Its immunoreactivity was examined by Western blotting. Results The recombinant expression plasmid was identified by PCR, double endonuclease digestion and sequencing. The recombinant Nus-P-29 was about Mr 93 000 with a concentration of 0.78 mg/ml. It was recognized by the sera of cystic echinococcosis patients. Conclusion The P-29 gene has been expressed with adequate immunoreactivity.

Key words: Echinococcus granulosus, Specific antigen gene, Prokaryotic expression