title: [Construction and expression of the major outer membrane protein PI of Neisseria gonorrhoeae in Escherichia coli].
abstract: OBJECTIVE
  To construct Neisseria gonorrhoeae major outer membrane protein PI gene recombinants for expression of the target protein in E. coli.
  
  
  METHODS
  Four clinic isolates of Neisseria gonorrhoeae were collected, and then the genome DNA of these strains was extracted. The gene encoding for PI of Neisseria gonorrhoeae was amplified by PCR, inserted into the cloning vector pBS-T; the recombinant plasmids pBS-T-PI-NG were constructed and sequence analysis was performed. Then PI gene fragments were inserted into expression vector pET30b to form pET30b-PI-NG recombinants. The PI protein expression was induced by adding IPTG in the inocula. The expressed proteins were analyzed by SDS-PAGE.
  
  
  RESULTS
  The pET30b-PI-NG expression recombinants for four clinic isolates of Neisseria gonorrhoeae were constructed successfully; three of the four expression recombinants were expressed successfully.
  
  
  CONCLUSIONS
  The expressed PI protein will be applied in the further research for PI antigenicity and immunological activity. This will be very helpful for the further construction of preventive vaccines directed against Neisseria gonorrhoeae infection.
authors.affiliation: Department of Medical Technology, West China School of Public Health, Sichuan University, Chengdu 610041, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Wen
authors.forename: Li
authors.initials: L
authors.lastname: Zhan
authors.forename: Dong-li
authors.initials: DL
authors.lastname: Wang
authors.forename: Xin
authors.initials: X
authors.lastname: Xu
authors.forename: Gang
authors.initials: G
authors.lastname: Yong
authors.forename: Xiao-fang
authors.initials: XF
authors.lastname: Pei
mesh_terms: D001428:Bacterial Vaccines / Q000096:biosynthesis / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D006801:Humans; D009344:Neisseria gonorrhoeae / Q000235:genetics* / Q000378:metabolism; D018272:Porins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
chemical_list: D001428:Bacterial Vaccines; D029968:Escherichia coli Proteins; D018272:Porins; D011993:Recombinant Fusion Proteins; C057699:porin protein, Neisseria