产气荚膜梭菌α毒素保护性抗原基因在大肠杆菌中的高效表达  

High level Expression of Alpha toxin Protective Antigen Gene of Clostridium perfringens in Escherichia coli

作　　者：许崇波[1] 朱平[1] 姚湘燕[1] 王卓[1] 冯书章[1] 马从林 孟锐奇[1] 

机构地区：[1]解放军农牧大学军事兽医研究所

出　　处：《中国兽医学报》1999年第1期29-32,共4页Chinese Journal of Veterinary Science

基　　金：全军医药卫生科研基金

摘　　要：对含有产气荚膜梭菌（Ｃｌｏｓｔｒｉｄｉｕｍｐｅｒｆｒｉｎｇｅｎｓ）α毒素基因的重组菌株ＢＬ２１（ＤＥ３）（ｐＸＥＴＡ１）和ＢＬ２１（ＤＥ３）ｐｌｙｓＳ（ｐＸＥＴＡ１），通过培养性状观察和小鼠接种试验，证明这２株重组菌株均无致病性。随后对这２株重组菌株的表达产物进行了研究，经ＳＤＳ－ＰＡＧＥ和薄层凝胶扫描分析，ＩＰＴＧ诱导３～４ｈ后的ＢＬ２１（ＤＥ３）（ｐＸＥＴＡ１）表达的α毒素占菌体总蛋白３３．２１％，ＢＬ２１（ＤＥ３）ｐｌｙｓＳ（ｐＸＥＴＡ１）表达的α毒素占菌体总蛋白２７．２５％，其相对分子质量约３７５００；经Ｗｅｓｔｅｒｎｂｌｏｔ分析，表达产物可被α毒素抗血清识别。包涵体粗提物的免疫攻毒试验结果表明，以１倍致死量攻击的免疫小鼠可获得１００％（３６／３６）的保护，以２倍致死量攻击的免疫小鼠可获得９４．２８％（３３／３５）的保护，从而说明表达产物具有良好的免疫原性。The alpha toxin antigen gene of Clostridium perfringens was expressed in recombinant strain BL21(DE3)(pXETA1) and BL21(DE3)plysS(pXETA1), and the expressed alpha toxin was recognized by anti sera of Clostridium perfringens type A with Western blot analysis. The two recombinant strains was proved to be unpathogenic. Immunization in a mouse model with crude preparation containing alpha toxin inclusion body induced protection against at least 2MLD of the toxin and Clostridium perfringens type A. The results showed that the expressed product of alpha toxin protective antigen gene possess a very good immunogenicity.

关 键 词：产气荚膜梭菌 Α毒素 基因表达 免疫原性 

分 类 号：S852.44[农业科学—基础兽医学] S855.12[农业科学—兽医学]