›› 2003, Vol. 21 ›› Issue (1): 6-19.

• 论著 • Previous Articles     Next Articles

Prokaryotic Expression and Characterization of an Antigenic Gene of Adult Trichinella spiralis

ZHU Xin-ping1,YANG Jing1,YANG Ya-ping1, Pascal Boireau2 ZHAN Bin3 FENG Jian-jun3,Peter Hotez3
   

  1. 1 Department of Parasitology,Capital University of Medical Sciences,Beijing 100054,China, 2 Umr Bipar Inra Afssa Enva Upvm,Maisons Alfort,France, 3 Department of Microbiology & Tropical Medicine,the George Washington University,Washington,DC,20037
  • Received:1900-01-01 Revised:1900-01-01 Online:2003-02-28 Published:2003-02-28

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Abstract: 　Objective To obtain a purified Ts87 gene expression product of adult Trichinella spiralis and identify its immunogenicity. Methods Ts87 cDNA was subcloned into PET-28a(+) expression vector. The transformed bacteria bearing PET-28a(+)/Ts87 plasmids were induced by IPTG for production of fusion proteins. The expression product purified with His binding affinity chromatography and electro elution assay was analyzed by SDS-PAGE, ELISA and Western blot, and was used to immunize the rabbits. Results PET-28a(+)/Ts87 transformed bacteria produced the desired fusion protein with a relative molecular weight of 40 kDa. The antisera with high titer were obtained by immunizing rabbit with Ts87 recombinant protein. Ts87 expression protein was detected as positive reaction with infected rabbit sera, infected swine sera and antisera against Ts87 by ELISA. Ts87 protein was also recognized by above-mentioned sera with Western blotting. However, Ts87 protein failed to react with the patient sera infected with Cysticercus cellulosae or Echinococcus granulosus, and rabbit sera infected with Schistosoma japonicum. Conclusion A new Trichinella recombinant protein with specific antigenicity was obtained.

Key words: Trichinella spiralis, gene expression, purification, recombinant protein, antigen, identification