title: [Construction and expression of recombinant plasmid pET28alpha-Sj26GST of Schistosoma japonicum in Escherichia coli BL21].
abstract: OBJECTIVE
  To construct and express recombinant plasmid pET28a-Sj26GST of Schistosoma japonicum (Sj) in Escherichia coli BL21 (DE3).
  
  
  METHODS
  The total RNA was extracted from Sj adult worms by ultrasound-breaking. The Sj26GST antigen gene was amplified by RT-PCR from the total RNA, and then cloned into prokaryotic expression plasmid pET28alpha and transformed into E. coli BL2 (DE3). The BL21(pET28a-Sj26GST) was induced with isopropyl-beta-D-thiogalactopyranosid (IPTG), and the expressed products were analyzed and identified with SDS-PAGE and Western blot.
  
  
  RESULTS
  The 676 bp Sj26GST gene was successfully amplified by RT-PCR and cloned into pET28alpha. The recombinant plasmid pET28a-Sj26GST was successfully constructed, with a relative molecular weight of expressed recombinant protein at approximately 36 x 10(3) as determined by SDS-PAGE. The amount of the expressed protein comprised 26% of the total bacterial proteins. The fusion protein could be recognized by the sera of rabbits infected with Sj.
  
  
  CONCLUSIONS
  The recombinant plasmid pET28alpha-Sj26GST is successfully constructed and highly expressed in E. coli in a fused form with His-tag. The expressed fusion protein shows specific antigenicity.
authors.affiliation: Institute of Infectious and Parasitic Diseases, the First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Wen-gui
authors.initials: WG
authors.lastname: Li
authors.forename: Bang-zhong
authors.initials: BZ
authors.lastname: Xiao
authors.forename: Xing-jian
authors.initials: XJ
authors.lastname: Luo
authors.forename: Ya-tang
authors.initials: YT
authors.lastname: Chen
authors.forename: Cheng-guo
authors.initials: CG
authors.lastname: Wu
mesh_terms: D000818:Animals; D000947:Antigens, Helminth / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D000939:Epitopes; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D010957:Plasmids / Q000096:biosynthesis / Q000235:genetics; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D012549:Schistosoma japonicum / Q000201:enzymology* / Q000235:genetics; D012554:Schistosomiasis japonica / Q000276:immunology; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology
chemical_list: D000947:Antigens, Helminth; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D005982:Glutathione Transferase