产气荚膜梭菌β2蛋白的高效可溶性表达与基因工程亚单位疫苗的制备

The efficient and soluble expression of β2 protein of Clostridium perfringens and the preparation of genetic engineering subunit vaccine

β2毒素是一种新近报道的产气荚膜梭菌（Clostridium perfringens）毒素,A、B、C、D、E等5个型的梭菌均可产生,该毒素也是制备基因工程亚单位疫苗的主要研究对象。本研究通过优化β2蛋白的密码子、去除蛋白信号肽,选择亲水性与抗原性较好的序列,同时优化表达条件等方法的探索,在大肠杆菌表达系统中获得了高效表达的可溶性重组β2蛋白,用该蛋白免疫小鼠可使动物产生较高的血清抗体水平,针对A型、B型、C型和D型产气荚膜梭菌的保护率分别为90%、100%、80%和90%,小鼠三免后第7~14天抗体效价达到峰值。本研究表达的β2蛋白具有较好的免疫原性,可进一步用于研制预防产气荚膜梭菌的基因工程亚单位疫苗。

β2 toxin is the most important toxin in all types of Clostridium perfringens（A,B,C,D and E）,and it is the main research object in preparing of genetic engineering subunit vaccine.According to optimization of codon,removal of the signal peptide,select better hydrophilicity and antigenic sequences,and optimize the expression conditions in this study,we obtained the soluble β2 fusion protein in Escherichia coli expression system.In addition,on the basis of β2 protein,we developed genetic engineering subunit vaccine for the inhibition of mixed infection of several types Clostridium perfringens.The serum antibody level increased obviously after immuning,and it could resist the attack of Clostridium perfringens.The vaccine could resist the attack of type A,B,C and D of Clostridium perfringens,and immune protection rate of the mouse were up to 90%,100%,80% and 90%,respectively.From day 7 to 14 after the third immune,the antibody titer in mice reached a peak. The vaccine can be used to control the disease caused by Clostridium perfringens infection,and it has a good application prospect.