猪传染性胃肠炎病毒S截短蛋白在干酪乳杆菌表面展示表达的研究  

Surface-displayed truncated spike glycoprotein of transmissible gastroenteritis virus on Lactobacillus casei

作　　者：田斌[1] 余丽芸[1,2] 侯喜林[2] 王桂华[1] 刘振格[2] 郭东伟[1] 刘秋晨[1] 齐浩[2] 林红丽[2] 

机构地区：[1]黑龙江八一农垦大学生命技术学院,黑龙江大庆163319 [2]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319

出　　处：《中国预防兽医学报》2013年第11期874-877,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省科技厅重点攻关课题(GB05B501-2)

摘　　要：为构建细菌外膜展示表达猪传染性胃肠炎病毒(TGEV)S截短蛋白的重组干酪乳杆菌(Lactobacillus casei),本研究采用RT-PCR扩增TGEV S基因的截短片段BC和AD,构建了重组pLA-BC/L.casei和pLA-AD/L.casei,利用western blot、间接免疫荧光法(IFA)和流式细胞术(FACS)检测外源蛋白在干酪乳杆菌的表面表达与定位。IFA和FACS结果显示重组菌株pLA-BC/L.casei和pLA-AD/L.casei表面均能够检测到截短的S和PgsA融合表达蛋白与TGEV阳性血清的特异性荧光信号,而对照菌没有检测到荧光信号。表明本研究构建的的TGEV重组L.casei具有胞外展示表达TGEV抗原蛋白的功能,可以作为TGEV候选疫苗应用于动物试验研究。To express the spike glycoprotein （S） of transmissible gastroenteritis virus （TGEV） on the surface of Lactobacillus casei, the BC and AD fragments of S gene were amplified by RT-PCR and inserted into the pLA shuttle vector （containing pgsA gene） to construct the recombinant plasmids of pLA-BC and pLA-AD, which were electroporated into L.casei to generate the recombinant bacteria pLA-BC/L.casei and pLA-AD/L.casei, respectively. In addition, The PgsA fusion proteins were expressed efficiently in the recombinant L.casei detected by western blot. Furthermore, IFA and FACS analysis showed that fluorescence signal was exhibited from both pLA-BC/L.casei and pLA-AD/L.casci, but not on control L.casei. The findings indicated that the TGEV S antigenic protein were surface-displayed on the recombinant L.casei and recombinant L.casei would be of a candidate TGEV vaccine and applied in fiLrther animal experiments.

关 键 词：猪传染性胃肠炎病毒 重组干酪乳杆菌 表面展示表达 

分 类 号：S852.65[农业科学—基础兽医学]