title: [Cloning and expression of the succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum Chinese strain in E. coli].
abstract: OBJECTIVE
  To clone the full-length gene encoding succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum (SjSDISP) Chinese strain and express it in Escherichia coli.
  
  
  METHODS
  According to the published incomplete EST (BU804141) of SjSDISP and the sequence of multiclone sites of lambda gt11 vector, 2 pairs of primers were designed and synthesized. Then the 3' and 5'ends of the EST of the SjSDISP from adult Schistosoma japonicum cDNA library were amplified by anchored PCR. After sequencing, a full-length cDNA sequence of the SjSDISP was obtained, and then it was cloned into prokaryotic expression vector pGEX-4T-1. Identified by agarosed gel electrophoresis, endonucleases digestion and PCR, the resultant recombinant plasmid was used for the expression under the temperature-dependent condition and Western blot analysis.
  
  
  RESULTS
  A 1,071 bp sequence was obtained. Sequence analysis showed that the fragment contained a complete open reading frame (ORF), encoding 278 amino acid residues. This target fragment was cloned into the prokaryotic expression vector pGEX-4T-1, and expressed in Escherichia coli. SDS-PAGE revealed that the molecular weight of the expressed fusion recombinant product was 56 kD. Western blot showed that the recombinant protein was recognized by polyclonal rabbit antiserum immunized with Schistosoma japonicum adult worm antigen.
  
  
  CONCLUSIONS
  Cloning of the full-length gene encoding SjSDISP and its bacterial expression were successfully done.
authors.affiliation: Department of Pathogenic Biology, Xiangya School of Medicine, Central South University, Changsha 410078, China.
authors.forename: Jun-long
authors.initials: JL
authors.lastname: Yu
authors.forename: Shi-ping
authors.initials: SP
authors.lastname: Wang
authors.forename: Zhuo
authors.initials: Z
authors.lastname: He
authors.forename: Gan
authors.initials: G
authors.lastname: Dai
authors.forename: Xiao-xin
authors.initials: XX
authors.lastname: Jiang
authors.forename: Shao-hua
authors.initials: SH
authors.lastname: Zeng
authors.forename: Xiao-qing
authors.initials: XQ
authors.lastname: Xiao
authors.forename: Song-hua
authors.initials: SH
authors.lastname: Zhou
authors.forename: Wen-kai
authors.initials: WK
authors.lastname: Li
authors.forename: Shao-rui
authors.initials: SR
authors.lastname: Xu
authors.forename: Zhi-yue
authors.initials: ZY
authors.lastname: Lü
authors.forename: Xian-chu
authors.initials: XC
authors.lastname: Peng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism*; D015801:Helminth Proteins / Q000096:biosynthesis / Q000235:genetics*; D007506:Iron-Sulfur Proteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012549:Schistosoma japonicum / Q000235:genetics / Q000378:metabolism*; D017385:Sequence Homology; D013385:Succinate Dehydrogenase / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D015801:Helminth Proteins; D007506:Iron-Sulfur Proteins; D011994:Recombinant Proteins; D013385:Succinate Dehydrogenase