口腔链球菌丙酮酸氧化酶基因缺陷型变异株的构建
Construction of the Pyruvate Oxidase Gene Def iciency Variant Strain of Streptococcus oralis
作者：庞若愚 张蓉 章锦才 张蕴惠 俞少杰
Author：Pang Ruoyu,Zhang Rong,Zhang Yunhui,et al
收稿日期：2002-02-25 年卷（期）页码：2002,20(01):62-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：口腔链球菌,丙酮酸氧化酶,同源重组,过氧化氢,变异,
Key words：Streptococcus oralis,pyruvate oxidase,homologous recombination,variation,
基金项目：
本课题为国家自然科学基金资助项目(编号39970793)
中文摘要
目的:利用口腔链球菌丙酮酸氧化酶基因(sopox) 的克隆序列,重组构建新的不能产生H2O2 的口腔链球菌变异株。方法:口腔链球菌株ATCC10557 经培养后用酚—氯仿法抽提细菌染色体基因组DNA ,经PCR 扩增sopox 基因,用BamHI 进行限制酶切;参照Chris 方法进行电转化,挑选阳性菌落测定其上清液中H2O2 的含量;将细菌传3～4 代后再次重复上述检测。结果:转化子经筛选后得到1 株阳性菌落,测定上清液中H2O2 含量,第1 次检测表明变异株产生H2O2 的量仅有所下降(介于阳性对照ATCC 10557 和阴性对照大肠杆菌JM109 之间) ,经3～4 次传代后变异株中上清液H2O2 量已经明显低于阴性对照。结论:成功构建了口腔链球菌丙酮酸氧化酶基因缺陷型变异株。
英文摘要
The purpose of this study is to construct a pyruvate oxidase gene deficiency variant strain of Streptococcus oralis ( S . oralis) . Methods : The sopox gene , which was got using polymerase chain reaction (PCR) , and the 130- basepair segment of which was cut down with endonuclease BamHI , and transferred into S . oralis (ATCC10557) by using electrotransformation. The authors obtained a variant strain of S . oralis , and then the catalase activity of the first culture and 3-4 subcultures was examined. Results : The authors obtained a pyruvate oxidase gene deficiency variant strain of S . orlis. The catalase activity examination showed that the ability of producing H2O2 of the variant strain of S . orlis declined , whose catalase activity was between those of the positive control (ATCC10557) and the negative control ( Escherichia coli , JM109) . But the produced H2O2 quantity of their subcultures was less than that of the negative control.Conclusion :The construction of the pyruvate oxidase gene deficiency variant strain of Streptococcus oralis is successful.
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