›› 2000, Vol. 18 ›› Issue (4): 8-223.

• 论著 • Previous Articles     Next Articles

GENE CLONING AND CHARACTERIZATION OF MITOCHONDRIA-RELATED PROTEIN OF SCHISTOSOMA JAPONICUM 

HU Xue-mei 2**;WU Hai-wei 1;ZHANG Zhao-song 1;SU Chuan 1;ZHAO wei 3;SHEN Lei 1;WANG Rong-zhi 1;MA Lei 1;ZHOU Ji-li 2;CHEN Shu-zhen 1;WU Guan-ling 1   

  1. 1 Research Laboratory of Molecular Inmmuoparasitology;Institute of Medical Molecular Biology,Nanjing Mwdical University,Nanjing 210029;2 Department of parasitology,Binzhou Medical College,Binzhou 256603;3 Department of Biology,Ningxia Medical College,Yinchuan 750004
  • Received:1900-01-01 Revised:1900-01-01 Online:2000-08-30 Published:2000-08-30

Abstract:  Objective] To subclone and characterize a cDNA clone coding for Schistosoma japonicum (S.j.) mitochondria-related protein. [Methods] The open reading frame of the fragment(Sj338/24) obtained from an adult worm cDNA library of S j. was analysed, at the upstream and downstream of the open reading frame(ORF) the primers A and B were designed,respectively, and the cDNA fragment was used as PCR template. The Sj338 gene fragment obtained was amplified by PCR method and then subcloned into pGEM-T vector for sequencing. The gene sequence was analyzed and the target fragment was restrictedly digested and subcloned into expression vector pGEX-6P-1. The expressed recombinant protein was purified and characterized. [Results] The cloned Sj338 gene was demonstrated to be 487 bp long containing one 459 bp ORF, encoding a protein with a molecular weight of 17 kDa.The nucleotide sequence of the cloned gene Sj338 had higher homology with those genes coding for mitochondrial outer membrane protein of Homo sapiens and Rattus norvegicus . The recombinant construct of pGEX-6P-1/Sj338 could be expressed efficiently and the antigenicity of its product rSj338 has been demonstrated by Western blotting. [Conclusion] Sj338 may be the gene coding for S j. mitochondria-related protein and the recombinant protein may be used as a new vaccine candidate .

Key words: Schistosoma japonicum, mitochondria, gene cloning, recombinant antigen, fusion expression