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

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(SOE-PCR)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80.16 kDa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 cells 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.