title: [Preparation and characteristics of Schistosoma japonicum signaling protein 14-3-3].
abstract: OBJECTIVE
  To prepare soluble recombinant signal transduction protein 14-3-3 of Schistosoma japonicum (rSj14-3-3) and investigate its immunologic features.
  
  
  METHODS
  The cDNA fragment of signal transduction protein 14-3-3 gene was prepared from Schistosoma japonicum adult worm mRNA, and was subcloned to the downstream of glutathione S transferase gene of expression vector pGEX-4T-3 to construct a recombinant expression plasmid 14-3-3/pGEX-4T-3. The recombinant plasmids were transferred into E. coli BL21, the transforments containing recombinant plasmid were induced by IPTG, and the expression situation of fusion protein GST-14-3-3 was observed by SDS-PAGE. The pure recombinant 14-3-3 protein was prepared by digesting the fusion protein GST-rSj14-3-3 with thrombin and affinity chromatography. The specific antibody against rSj14-3-3 protein was prepared by an immunized rabbit with rSj14-3-3 protein. The immunogenicity and immune reactivity of rSj14-3-3 protein were analyzed by Western blotting.
  
  
  RESULTS
  The gene encoding signal transduction protein 14-3-3 of Schistosoma japonicum Jiangsu strain was cloned successfully, the homology of open read frame sequence to the registrated sequence of Sj14-3-3 protein in genbank was 99.08%. A 55 kilo Dalton fusion protein GST-rSj14-3-3 was expressed by transferring the recombinant plasmid 14-3-3/pGEX-4T-3 into E. coli BL21 and induced with IPTG. The high titer antibody against rSj14-3-3 was produced by the immunized rabbit with rSj14-3-3 ,and could recognize the nature and recombinant rSj14-3-3 proteins.
  
  
  CONCLUSIONS
  The soluble rSj14-3-3 protein has been prepared successfully in this study, and it has good immunogenicity and reactivity.
authors.affiliation: Jiangsu Institute of Parasitic Diseases, Key Laboratory on Technology for Parasitic Disease Prevention and Control, Ministry of Health, Wuxi 214064, China.
authors.forename: Xu-Ren
authors.initials: XR
authors.lastname: Yin
authors.forename: Chuan-Xin
authors.initials: CX
authors.lastname: Yu
authors.forename: Shu-Ying
authors.initials: SY
authors.lastname: Xie
authors.forename: Chun-Yan
authors.initials: CY
authors.lastname: Qian
authors.forename: Li-Jun
authors.initials: LJ
authors.lastname: Song
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Yong-liang
authors.initials: YL
authors.lastname: Xu
mesh_terms: D048948:14-3-3 Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D000818:Animals; D003001:Cloning, Molecular; D015870:Gene Expression; D015801:Helminth Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D008970:Molecular Weight; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012549:Schistosoma japonicum / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D015398:Signal Transduction
chemical_list: D048948:14-3-3 Proteins; D015801:Helminth Proteins; D011994:Recombinant Proteins