[Construction and expression of Neisseria surface protein (nspA) of Neisseria gonorrhoeae in Escherichia coli].
OBJECTIVE
To construct neisseria surface protein (NspA) recombinants of Neisseria gonorrhoeae from a reference strain and express this protein in E. coli.


METHODS
The fragments of NspA gene of Neisseria gonorrhoeae was amplified by PCR from the reference strain genomic DNA and cloned into expression vector pET-30c (+) to get the pET-NspA recombinants. The recombinants were verified with restrictive endonuclease digestion and sequence analysis. The verified recombinant was transformed into E. coli BL21 (DE3). After inducing with IPTG, the expressed NspA protein was analyzed by SDS-PAGE and Western Blot.


RESULTS
The pET-NspA expression recombinants for the reference strain of Neisseria gonorrhoeae were successfully constructed and the induced recombinant NspA protein was observed.


CONCLUSIONS
The successful expression of the Neisseria gonorrhoeae NspA protein will be very helpful for the further research of its antigenicity and immunological activity, and for the construction of preventive vaccines on Neisseria gonorrhoeae infection.
Zhan
Li
L
Department of Medical Technology, West China School of Public Health, Sichuan University, Chengdu 610041, China.
Zhang
Chao-Wu
CW
Wang
Dong-Li
DL
Xu
Xin
X
Meng
Ling
L
He
Chao
C
Zhou
Ai-Ping
AP
Pei
Xiao-Fang
XF
D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D009344:Neisseria gonorrhoeae / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D001425:Bacterial Outer Membrane Proteins; C105506:NspA protein, Neisseria; D011994:Recombinant Proteins