串联IL-2的犬瘟热病毒F-H融合基因重组乳酸杆菌的构建与表达  

Construction and Expression of Recombinant Lactobacillus Containing IL2-F-H Fusion Gene of Canine Distemper Virus

作　　者：苏凤艳[1] 李哲[1] 李艳芝 王春凤 曾范利[1] SUFeng-yan LI Zhe LI Yan-zhi WANG Cun-feng ZENG Fan-li(College of Chinese Medicinal Materials,Jilin Agricultural University,Changchun 130118, China Research Center of Animal Probiotics Engineering in Jilin Province, Changchun 130118, China Animal Husbandry Work Station of Yitong Manzhu Autonomous County in Jilin, Yitong 130700, China)

机构地区：[1]吉林农业大学中药材学院,长春130118 [2]吉林省伊通满族自治县畜牧工作总站,吉林伊通130700 [3]吉林省动物微生态制剂工程研究中心,长春130118

出　　处：《上海交通大学学报（农业科学版）》2017年第4期83-88,共6页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：吉林省重点科技攻关项目(20130206039NY)

摘　　要：为了研发犬瘟热乳酸杆菌载体疫苗,更好地预防犬瘟热,试验采用重叠延伸PCR(SOEPCR)技术扩增IL2-F-H融合基因,将IL2-F-H融合基因亚克隆至穿梭载体pSIP409上,构建pSIP-IL2-F-H重组表达载体,并电转化至植物乳杆菌NC8感受态细胞中,构建串联水貂白细胞介素-2(IL-2)的犬瘟热病毒(CDV)F-H融合基因重组乳酸杆菌;利用SDS-PAGE和Western-blot检测IL2-F-H融合蛋白的表达情况。检测表明,重组乳酸杆菌表达了分子量约为80.16kDa的IL2-F-H融合蛋白,且该融合蛋白能与CDV抗体发生反应,具有反应原性。In order to study and develop the Lactobacilli carrier vaccine of canine distemper, the IL2-F-H fusion gene was amplified by gene splicing by overlap extension PCR （SOE-PCR）. The IL2-F-H fusion gene was sub-cloned into shuttle carrier pSIP409 to construct a recombinant plasmid （named as pSIP-IL2- F-H）. Then the recombinant plasmid was transformed into Lactobacillus plantarum competence celis by electroporation. Recombinant Lactobacillus containing a IL2-F-H fusion gene of canine distemper virus （CDV） was constructed. SDS-PAGE and Western-blot were used to detect the expression of IL2-F-H fusion gene. Results showed that the recombinant Lactobacillus expressed a fusion protein （nearly 80. 16 kDa in size） which reacted with CDV antibody.

关 键 词：犬瘟热病毒 IL2-F-H融合基因 乳酸杆菌 表达 

分 类 号：S852.65[农业科学—基础兽医学]