The dominant genotypes of porinⅠ gene of Neisseria gonorrhoeae isolates in Zhejiang Province and the construction of the prokaryotic expression system / 预防医学

ABSTRACT

Objective To analyze the dominant genotypes of porin Ⅰ gene encoding major outer membrane protein of Neisseria gonorrhoeae in Zhejiang Province and to construct the prokaryotic expression systems of porinⅠA and porinⅠB genes.Methods Based on the previous research,the porinⅠA and porinⅠB genes were sequenced after T -A cloning, and the dominant genotypes of porinⅠA and porinⅠB genes were analyzed.Then the prokaryotic expression systems of the dominant genotypes of porinⅠA and porinⅠB genes were constructed.Ten percent SDS -PAGE was applied to measure the output of PⅠA and PⅠB proteins after inducement by 0.5 mmol/L IPTG.Results All 5 porinⅠA isolates sequenced were serovar ⅠA -6.Of the 11 porinⅠB isolates sequenced,there were 5 serovarⅠB -3 isolates,3 serovarⅠB -3 /6 isolates,1 serovarⅠ B -6 isolate and 2 mutant isolates.Outputs of P Ⅰ A and P Ⅰ B expressed by the constructed prokaryotic expression systems PET -42 -PⅠA and PET -42 -PⅠB were as high as 30% and 20% respectively. Conclusion ⅠA -6 is the dominant genotypes of porinⅠA gene and ⅠB -3 is the dominant genotypes of porinⅠB gene in Zhejiang Province.Comparing to the porinⅠB gene,porinⅠA gene is more conserved.The prokaryotic expression systems with high efficiency of porinⅠA and porinⅠB genes were successfully constructed,which may be helpful in the further research of genetic engineering vaccine and clinical detection of Neisseria gonorrhoeae.