表达鸡柔嫩艾美耳球虫表面抗原TATA4与鸡IL-IL-2的侵入型乳酸菌构建及鉴定  

Construction and Identification of Invasive Lactic Acid Bacteria with E. tenella Surface Antigen TA4 and Chicken IL-2

作　　者：刘洋[1,2,3] 姜延龙 刘晶[1,2,3] 张赞[1,2,3] 高兴 王春凤[1,2,3] 杨桂连 LIU Yang;JIANG Yanlong;LIU Jing;ZHANG Zan;GAO Xing;WANG Chunfeng;YANG Guilian(College of Animal Science and Technology,Changchun,Jilin 130118;Jilin Provincial Engineering Research Center of Animal Probiotics,Changchun,Jilin 130118;Key Laboratory of Animal Production and Quality Security,Ministry of Education,Changchun,Jilin 130118)

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118 [2]吉林省动物微生态制剂工程研究中心,吉林长春130118 [3]动物生产及产品质量安全教育部重点实验室,吉林长春130118

出　　处：《中国家禽》2019年第10期20-25,共6页China Poultry

基　　金：国家重点研发计划项目(2017YFD0501200);国家自然科学基金项目(31672528、31602092);吉林省科技发展计划项目(20160519011JH、20170204034NY、20180201040NY);吉林省产业创新专项资金项目(2016C063)

摘　　要：为探究鸡柔嫩艾美耳球虫(E. tenella)表面抗原TA4在植物乳杆菌NC8的表达情况,使用PCR扩增TA4片段,同时使用细胞因子ChIL-2作为免疫佐剂,应用猪圆环病毒2A肽(P2A)序列使抗原与免疫佐剂共表达,连接有金黄色葡萄球菌的纤连蛋白结合蛋白A(FnBPA)的409载体,构建两株重组侵入型乳酸菌NC8 pSIP 409-FnBPA-pgsA′-TA4和NC8 pSIP 409-FnBPA-pgsA′-TA4-IL2。使用自制多克隆抗体通过Western blot检测重组菌的蛋白表达情况。结果表明,两种侵入型目的蛋白TA4和IL-2使用多克隆抗体检测均可成功检测到蛋白表达,为后期动物试验及益生菌口服疫苗的制备奠定了基础。In order to investigate the expression of E.tenella surface antigen TA4 in Lactobacillus plantarum NC8,the TA4 fragment was amplified by PCR,and the cytokine ChIL-2 was used as an immunoadjuvant.The virus 2 A peptide(P2 A)sequence co-expressed the antigen with an immunoadjuvant,and 409 vector of fibronectin-binding protein A(FnBPA)linked to S.aureus was constructed two recombinant invasive lactic acid bacteria NC8 pSIP 409-FnBPApgsA′-TA4 and NC8 pSIP 409-FnBPA-pgsA′-TA4-IL2.The protein expression of the recombinant strain was detected by Western blot using a self-made polyclonal antibody.The results showed that the two target proteins,TA4 and IL-2,were successfully detected by polyclonal antibody detection,which laid a foundation for the animal experiments and the preparation of probiotic oral vaccine.

关 键 词：侵入型乳酸菌 鸡柔嫩艾美耳球虫 TA4 ChIL-2 

分 类 号：S852.723[农业科学—基础兽医学]