title: [Expression and characterization of recombinant Sj23HD-HSA fusion protein in Saccharomyces cerevisiae].
abstract: OBJECTIVE
  To prepare the fusion protein of large hydrophilic domain of 23 kDa membrane protein of Schistosoma japonicum and the mature peptide of human serum albumin (Sj23HD-HSA) and investigate its immunoreactivity.
  
  
  METHODS
  A fusion protein gene encoding Sj23HD-HSA fusion protein was prepared by overlapping PCR, which was confirmed by TA cloning and DNA sequencing. The fusion gene of Sj23HD-HSA was directionally subcloned into yeast expression plasmid pWX530 to construct a recombinant plasmid Sj23HD-HSA/pWX530. The transformants of Saccharomyces cerevisiae containing the recombinant plasmid Sj23HD-HSA/pWX530 were screened on leu deficient SD medium after yeast competent cells were transformed with recombinant plasmid. The excretive Sj23HD-HSA protein was expressed by culturing the yeast transformants at 30 degrees C for 1 week, and the protein component of culture supernatant was analyzed by SDS-PAGE. Sj23HD-HSA fusion protein was purified through Ion Exchange Chromatography. The immunoreactivity of recombinant Sj23HD-HSA fusion protein was determined by Western blotting with sera of schistosomiasis, clonorchiasis and healthy.
  
  
  RESULTS
  The gene encoding the Sj23HD-HSA fusion protein was constructed successfully, which was confirmed by DNA sequencing. The yeast transformants containing plasmid Sj23HD-HSA/pWX530 could express the excretive Sj23HD-HSA fusion protein without inducing. The results of Western blotting indicated Sj23HD-HSA could be recognized by the sera of schistosomiasis, but could not be recognized by the sera of clonorchiasis and healthy respectively.
  
  
  CONCLUSIONS
  Sj23HD-HSA fusion protein with good immune reactivity is prepared successfully, which will be a potential antigen for schistosomiasis immunodiagnosis.
authors.affiliation: Department of Novel Biotechnique, Jiangsu Institute of Parasitic Diseases, Key Laboratory on Technology for Parasitic Diseases Prevention and Control, Ministry of Health, Wuxi 214064, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Chuan-Xin
authors.initials: CX
authors.lastname: Yu
authors.forename: Jian-Linag
authors.initials: JL
authors.lastname: Yang
authors.forename: Wei
authors.initials: W
authors.lastname: Feng
authors.forename: Xu-Ren
authors.initials: XR
authors.lastname: Yin
authors.forename: Li-Jun
authors.initials: LJ
authors.lastname: Song
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
authors.forename: Chun-Yan
authors.initials: CY
authors.lastname: Qian
authors.forename: Xue-Dan
authors.initials: XD
authors.lastname: Ke
mesh_terms: D000818:Animals; D015870:Gene Expression; D015801:Helminth Proteins / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D012549:Schistosoma japonicum / Q000235:genetics / Q000276:immunology; D012554:Schistosomiasis japonica / Q000276:immunology* / Q000469:parasitology; D012709:Serum Albumin / Q000235:genetics / Q000276:immunology*
chemical_list: D015801:Helminth Proteins; D011993:Recombinant Fusion Proteins; D012709:Serum Albumin