口腔链球菌丙酮酸氧化酶基因调节区的DNA序列分析

华西口腔医学杂志

口腔链球菌丙酮酸氧化酶基因调节区的DNA序列分析

章锦才,张蓉,张蕴惠

510280　广东省口腔医院牙周科(章锦才),四川大学华西口腔医院牙周科(张　蓉,张蕴惠)

收稿日期:2003-10-25 修回日期:2003-10-25 出版日期:2003-10-20 发布日期:2003-10-20
基金资助:
本课题为国家自然科学基金资助项目(编号39970793)

DNA Sequence Analysis for the Promoter of Pyruvate Oxidase Gene fromStrptococcus Oralis

ZHANG Jincai*,ZHANG Rong,ZHANG Yunhui

*Department ofPeriodontology,Guangdong Provincial StomatologicalHospi- tal,Guangzhou510280,China

Received:2003-10-25 Revised:2003-10-25 Online:2003-10-20 Published:2003-10-20

摘要/Abstract

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1·30 kb片段和PBK-CMV质粒DNA用 HindⅢ单酶切产物连接,转化E.coliJM109,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1·30 kb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1 350 bp。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。

关键词: 口腔链球菌, 丙酮酸氧化酶, 丙酮酸氧化酶基因, 调节基因

Abstract:

Objective　To elucidate the molecular structure of pyruvate oxidase gene promoter.Methods　The 1·30 kb fragment with promoter activity, amplified from upstream ofStreptococcus oralispyruvate oxidase gene (Sopox), was cloned into vector PBK-CMV. The positive transformedE.coliJM109 clone was selected, the recombinant plasmidwas further identifiedwith restriction mapping analysis. The positive recombinant plasmid was studied with sequence analysis.Results　After digesting the recombinant plasmid withHindⅢ, 1% agarose electrophoresis showed 1·30 kb fragment, which was consistent with predicted size. Sequence analysis revealed 1 350 bp.Conclusion　TheSopoxpromoter region is sequenced. Further characterization of the Sopoxpromoter region will elucidate the molecular mechanism of H2O2production ofstreptococcus oralis.

Key words: Streptococcus oralis, pyruvate oxidase, pyruvate oxidase gene, promoter

章锦才,张蓉,张蕴惠. 口腔链球菌丙酮酸氧化酶基因调节区的DNA序列分析[J]. 华西口腔医学杂志.

ZHANG Jincai*,ZHANG Rong,ZHANG Yunhui. DNA Sequence Analysis for the Promoter of Pyruvate Oxidase Gene fromStrptococcus Oralis[J]. West China Journal of Stomatology.

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