[Construction and expression of Neisseria surface protein (nspA) of Neisseria gonorrhoeae in Escherichia coli].

Sichuan Da Xue Xue Bao Yi Xue Ban

Department of Medical Technology, West China School of Public Health, Sichuan University, Chengdu 610041, China.

Published: March 2006

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.

Conclusion: 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.

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