[Construction and expression of the major outer membrane protein PI of Neisseria gonorrhoeae in Escherichia coli].
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.
Wen
Hua
H
Department of Medical Technology, West China School of Public Health, Sichuan University, Chengdu 610041, China.
Zhan
Li
L
Wang
Dong-li
DL
Xu
Xin
X
Yong
Gang
G
Pei
Xiao-fang
XF
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
D001428:Bacterial Vaccines; D029968:Escherichia coli Proteins; D018272:Porins; D011993:Recombinant Fusion Proteins; C057699:porin protein, Neisseria