血链球菌丙酮酸氧化酶基因表达克隆的构建与鉴定

Expression of Pyruvate Oxidase Gene Sopox from Streptococcus Sanguis in E. coli

目的 构建血链球菌丙酮酸氧化酶基因 Sopox的表达质粒 ,为研究丙酮酸氧化酶基因 Sopox表达的调节奠定基础。方法 将从血链球菌 ATCC10 5 5 7克隆的丙酮酸氧化酶基因 Sopox重组到 p BV2 2 0 ,表达载体 ,并转化入大肠杆菌 E.coli JM10 5 ,观察 Sopox基因在大肠杆菌中的表达。结果 通过酶切后电泳鉴定 ,Sopox基因重组入 p BV2 2 0并转化入 E.coli JM10 5 ;经 42℃诱导后 ,SDS- PAGE显示 p BV2 2 0 / Sopox/ JM10 5表达分子量约为6 5 kd的蛋白 ,与根据 Sopox基因 DNA序列预测的蛋白分子量一致 ;p BV2 2 0 / Sopox/ JM10 5的蛋白表达在 42℃诱导 4小时后达高峰。结论 丙酮酸氧化酶基因 Sopox已被成功地克隆到 p BV2 2 0 ,并在 E.coli JM10

Objective To reconstruct the expression plasmid o f Sopox gene for further understanding the regulation of its expression. Methods Sopox was recombined with expression vector pBV220 and the ex pression of Sopox in E. coli JM105 was observed after transformation. Re sults pBV220/Sopox/JM105 expressed a protein with molecular weight of 6 5 kd on SDS PAGE after induction, and the expression reached the maximal amount with induction at 42℃ for 4 hours. Conclusion Sopox was succes sfully cloned into pBV220 and expressed in E.coli JM105.