乳糖诱导剂对重组C型产气荚膜梭菌α毒素基因表达的影响

Influences of lactose as an inducer on the expression of the alpha toxin gene of Clostridium perfringens type C

采用限制性核酸内切酶酶切鉴定含α毒素基因的重组质粒,用SDS-PAGE检测不同条件下α毒素基因的表达情况。经酶切鉴定证实重组质粒pXETA02含有α毒素基因且基因序列和阅读框架正确。结果表明,以乳糖诱导α毒素基因表达的优化条件为:培养基pH 7.0,培养温度37℃,菌体生长密度D600达到1.0时分批添加0.5g/L乳糖,诱导5h,此时目的蛋白表达量为23%,实现了α毒素基因的高效表达。从而为C型产气荚膜梭菌α毒素基因工程菌苗的生产工艺研究提供了可靠的试验数据。

To identify the recombinant plasmid pXETA02 with restriction endonucleases digestion and detect expres- sional level of alpha toxin gene under different conditional by SDS-PAGE. By identification of endonucleases digestion,the recombinant expression plasmid pXETA02contained alpha toxin gene with positive reading frame. The lactose was adopted as inducer in this experiment and the recombinant strain BL21（DE3） （pXETA02） were optimized by changing pH of medium,temperature,amount of inducer and inducing time. This provided experimental data to research productive technique of the genetically engineered vaccine of the alpha toxin gene of Clostridiurn perfringens type C.