不同诱导剂及培养条件对C型产气荚膜梭菌β_2毒素基因表达的影响  

Effect of different inducers and culture conditions on the expression of β_2-toxin gene from 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：《浙江农业学报》2015年第10期1822-1827,共6页Acta Agriculturae Zhejiangensis

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

摘　　要：对产气荚膜梭菌(Clostridium perfringens)β2毒素基因的表达载体p ETXB2进行限制性核酸内切酶酶切鉴定,并用SDS-PAGE检测了不同培养条件下,该毒素基因在埃希氏大肠杆菌(Escherichia coli)BL21(DE3)工程菌株中的表达情况。结果表明,p ETXB2重组质粒中的β2毒素基因位于p ET-28c(+)表达盒中,且阅读框架和基因序列正确。以IPTG为诱导剂时,重组菌株BL21(DE3)(p ETXB2)的最佳培养条件为37℃,p H7.0,IPTG终浓度0.8 mmol·L-1,诱导4 h;以乳糖为诱导剂时,其最佳培养条件为37℃,p H 7.0,乳糖终浓度0.5 g·L-1,诱导5 h。Recombinant plasmid p ETXB2 which contained Clostridium perfringens β2-toxin gene was identified with restriction endonucleases digestion,and the gene＇s expression products in Escherichia coli strain BL21（ DE3） under different culture conditions were detected by SDS-PAGE. The results were as follows： Endonucleases digestion and DNA sequencing confirmed that the recombinant expression plasmid p ETXB2 contained the β2-toxin gene in the expression cassette of p ET-28c（ ＋） with a positive reading frame. The optimization condition of inducible expression for β2-toxin by IPTG was 0. 8 mmol·L- 1of IPTG concentration and inducing for 4 h at 37℃,p H 7. 0. When induced by lactose,the optimization condition was 0. 5 g·L- 1of lactose concentration and inducing for 5 h at 37℃,p H 7. 0.

关 键 词：C型产气荚膜梭菌 β2毒素 IPTG 乳糖 

分 类 号：S852.61[农业科学—基础兽医学]