口腔链球菌丙酮酸氧化酶基因调节区的DNA序列分析  

DNA Sequence Analysis for the Promoter of Pyruvate Oxidase Gene from Strptococcus Oralis

作　　者：章锦才[1] 张蓉[2] 张蕴惠[2] 

机构地区：[1]广东省口腔医院牙周科,510280 [2]四川大学华西口腔医院牙周科

出　　处：《华西口腔医学杂志》2003年第5期350-352,共3页West China Journal of Stomatology

基　　金：国家自然科学基金资助项目 (编号 39970 793)

摘　　要：目的　了解口腔链球菌丙酮酸氧化酶基因调节区的分子结构。方法　将从口腔链球菌丙酮酸氧化酶基因 (Sopox)的上游区序列体外扩增得到的 ,并经证明含有启动子活性的 1 30kb片段和PBK_CMV质粒DNA用HindⅢ单酶切产物连接 ,转化E .coliJM10 9,筛选阳性菌落 ,增菌后取质粒DNA ,再经HindⅢ酶切鉴定。将阳性克隆进行DNA序列分析。结果　重组克隆酶切产物经 1%琼脂糖凝胶电泳显示 ,获得约为 1 30kb的条带 ,与预计片段大小相符证实其为阳性重组克隆。重组克隆测序获得Sopox基因上游序列共计 135 0bp。结论　Sopox基因调节区序列的获得为进一步阐明口腔链球菌产生过氧化氢的分子机理奠定基础。Objective\ To elucidate the molecular structure of pyruvate oxidase gene promoter.Methods\ The 1 30 kb fragment with promoter activity, amplified from upstream of Streptococcus oralis pyruvate oxidase gene (Sopox), was cloned into vector PBK_CMV. The positive transformed E.coli JM109 clone was selected, the recombinant plasmid was further identified with restriction mapping analysis. The positive recombinant plasmid was studied with sequence analysis.Results\ After digesting the recombinant plasmid with HindⅢ, 1% agarose electrophoresis showed 1 30 kb fragment, which was consistent with predicted size. Sequence analysis revealed 1 350 bp.Conclusion\ The Sopox promoter region is sequenced. Further characterization of the Sopox promoter region will elucidate the molecular mechanism of H\-2O\-2 production of streptococcus oralis.

关 键 词：口腔链球菌 丙酮酸氧化酶 基因调节 DNA序列 真核表达载体 克隆 

分 类 号：R780.2[医药卫生—口腔医学]