表面展示C型产气荚膜梭菌α，β毒素蛋白重组乳酸菌的构建及反应原性分析  

Construction of Recombinant Lactic Acid Bacteria with Surface Displayed Clostridium perfringen α and β Toxin Proteins and the Reactinogenicity Analysis

作　　者：李超[1] 刘孟宇 刘琼[2] 王春凤[2] 倪宏波[1] Li Chao;Liu Mengyu;Liu Qiong;Wang Chunfeng;Ni Hongbo(College of Animal Science and Technology,Heilongjiang Bayi Agricultural University,Daqing 163319;College of Animal Science and Technology,Jilin Agricultural University)

机构地区：[1]黑龙江八一农垦大学动物科技学院,大庆163319 [2]吉林农业大学动物科技学院

出　　处：《黑龙江八一农垦大学学报》2019年第6期23-28,共6页journal of heilongjiang bayi agricultural university

基　　金：国家重点研发计划(2017YFD0501000)

摘　　要：构建表达C型产气荚膜梭菌α,β1和β2毒素蛋白的重组乳酸菌,并对表达蛋白反应原性进行鉴定。通过PCR构建pSIP409-pgsA’-α,pSIP409-pgsA’-β1以及pSIP409-pgsA’-β2质粒,并将三种质粒电转入植物乳杆菌NC8后诱导表达,通过Western-blot和间接免疫荧光检测目的蛋白的表达情况。构建的NC8-pSIP409-pgsA’-α,NC8-pSIP409-pgsA’-β1以及NC8-p SIP 409-pgsA’-β2三种重组乳酸菌能成功表达C型产气荚膜梭菌α,β1和β2毒素蛋白,三种表达的融合蛋白大小分别为61 kDa、52 kDa以及46 kDa。三种重组乳酸菌表达了C型产气荚膜梭菌α,β1和β2毒素蛋白且与制备的多抗具有良好的反应原性,为C型产气荚膜梭菌口服疫苗的研制奠定基础。To construct recombinant lactic acid bacteria expressing Clostridium perfringen α,β1 and β2 proteins and identify its reactinogenicity,the gene was amplified from the DNA of Clostridium perfringen type c(cvcc1158)strain by PCR and inserted into anchoring vector pSIP409-pgsA’,Construction of pSIP409-pgsA’-α,pSIP409-pgsA’-β1 and pSIP409-pgsA’-β2 plasmids. After transformation plasmids into Lactobacillus plantarum NC8 strain,the synthesis of protein was determined by Western-blot and indirect immunofluorescence assay. The newly functional recombinant lactic acid bacteria could successfully express Clostridium perfringens type C α,β toxin proteins. The expressed proteins were identified by Western-blot and Indirect immunofluorescence with reactinogenicity. The expressed protein had good reactinogenicity,which provided a basis for later experiments.

关 键 词：pSIP409-pgsA’ C型产气荚膜梭菌 重组乳酸菌 反应原性 

分 类 号：S852.61[农业科学—基础兽医学]