淋病奈瑟菌外膜蛋白PorB编码基因的克隆及序列分析

目的 获取淋病奈瑟菌外膜蛋白PorB编码基因(porB)并构建其重组表达质粒。方法 根据porB已知序列，设计合成一对引物，用PCR方法从淋病奈瑟菌基因组DNA中扩增出porB基因片段，克隆到原核表达质粒pQE30中，转化大肠埃希菌M15感受态细胞，经酶切和PCR鉴定，然后进行测序。结果 porB基因体外扩增产物大小约为911bp。重组质粒经双酶切和PCR鉴定表明为正确重组子，测序结果与已知序列基本吻合。结论 成功克隆了淋病奈瑟菌外膜蛋白PorB编码基因，为下一步PorB蛋白的功能研究和淋病奈瑟菌蛋白质疫苗的研制提供了基本的物质基础：

Cloning and Sequence Analysis of Porin PorB Encoding Gene of Neisseria Gonorrhoeae

Objective To obtain DNA of Neisseria gonorrhoeae porin PorB, and construct a recombinant plasmid containing gene encoding PorB for nucleotide sequence analysis. Methods A couple of primers were designed for PCR according to the known sequence of porB. The porB gene was amplified by PCR from genome of Neisseria gonorrhoeae WHOE stain and cloned into plasmid pQE30. The recombinant plasmid pQE30-porB was transferred into competent E.coli M15. The recombinants were screened and identified by restriction analysis and PCR, the cloned gene was sequenced . Results The size of amplified porB gene was 911 bp. The correct recombinant plasmid pQE30-porB was isolated and confirmed by restriction analysis and PCR. DNA sequencing showed the DNA sequence of the cloned gene was the same as the published sequence. Conclusion The porB gene was successfully amplified and cloned into plasmid pQE30. It provided the basic material for studying the function of Porin PorB and the development of Neisseria gonorrhoeae protein vaccine.