IPTG诱导剂对C型产气荚膜梭菌β_2毒素基因表达的影响  

Influences of IPTG Inducer on Expression of the β_2-toxin Gene of Clostridium perfringens Type C

作　　者：许崇利[1] 

机构地区：[1]吉林化工学院生物与食品工程学院,吉林吉林132022

出　　处：《湖北农业科学》2016年第3期749-753,共5页Hubei Agricultural Sciences

基　　金：国家自然科学基金项目(30972188);吉林化工学院博士启动基金项目

摘　　要：对已构建的含β2毒素基因重组质粒p ETXB2进行限制性核酸内切酶酶切鉴定,并用SDS-PAGE检测不同培养条件下β2毒素基因的表达情况。经酶切鉴定证实,p ETXB2重组质粒含有β2毒素基因而且阅读框架和基因序列正确。同时采用IPTG诱导剂在改变培养基的p H、培养温度、诱导剂的加入量及诱导时间4个方面对β2毒素基因表达进行调控,其最佳的表达条件是37℃,p H 7.0的培养基中,用终浓度为0.8 mmol/L IPTG诱导4 h,β2毒素蛋白的表达效果最好。An identification of recombinant plasmid p ETXB2 containing β2-toxin gene was conducted with restriction endonuclease enzyme digestion, and the expression levels of β2-toxin gene under different cultural conditions were detected.By enzyme digestion identification, the recombinant plasmid p ETXB2 contained β2-toxin gene and have positive reading frame. Meanwhile IPTG was adopted as inducer to regulate and control expression of the β2-toxin gene in four aspects of changing medium p H, culture temperature, amount of inducer, and inducing time. Results showed that, the optimal condition was the temperature 18 ℃, medium p H 7.0, final concentration 0.8 mmol / L IPTG, inducing time 4 hours, the expression effect of β2-toxin protein was optimized under the condition.

关 键 词：C型产气荚膜梭菌 β2毒素基因 优化表达 

分 类 号：S852[农业科学—基础兽医学] Q78[农业科学—兽医学]