产气荚膜梭菌ε毒素重组突变体大肠杆菌表达条件的优化

Optimization of expression conditions for recombinant mutants of Clostridium perfringens ε-toxin in E.coli

关键词:  产气荚膜梭菌  ε毒素  基因工程亚单位疫苗  蛋白表达  

Key words:  Clostridium perfringens  ε toxin  genetic engineering subunit vaccine  protein expression  

摘要:

本研究旨在优化产气荚膜梭菌ε毒素重组蛋白的表达条件，为下一步大规模生产ε毒素基因工程亚单位疫苗奠定基础。以本实验室构建的产气荚膜梭菌ε毒素重组突变体为基础，通过控制变量法确定诱导温度、诱导时间、IPTG诱导浓度以及诱导时菌液浓度等原核蛋白表达条件。结果表明，ε毒素的重组突变体在37 ℃、菌液OD600值为0.895~1.295时，以1.6 mM IPTG诱导6 h条件下可溶性表达量最高。

Abstract:

：To optimize the expression conditions of the ε-toxin recombinant protein of Clostridium perfringens and lay the foundation for the next large-scale production of ε-toxin genetic engineering subunit vaccines. Based on the recombinant mutant of Clostridium perfringens epsilon toxin constructed in our laboratory, the prokaryotic protein expression conditions such as induction temperature, induction time, IPTG induction concentration and bacterial solution concentration during induction were determined by the control variable method. The results showed that the recombinant mutant of epsilon toxin had the highest soluble expression under the condition of 6 h induced by 1.6 mM IPTG at 37 ℃ and the OD600 value of the bacterial solution was 0.895~1.295.