O型口蹄疫病毒VP1T细胞、B细胞表位双拷贝基因与大肠杆菌LTB基因融合表达产物的免疫应答  

Study on the Immune Response of a Fusion Expression Product Consisted of Two-copy Gene of T-cell and B-cell Epitopes of Type O of Food-and-mouth Disease Virus VP1 and LTB Gene of Escherichia coli

作　　者：庄娟[1] 尤永进[2] 陈波[2] 饶忠[2] 潘洁[2] 

机构地区：[1]淮阴师范学院生物系,江苏淮安223001 [2]上海农业科学院畜牧兽医研究所,上海201106

出　　处：《安徽农业科学》2007年第33期10622-10623,10631,共3页Journal of Anhui Agricultural Sciences

摘　　要：[目的]为预防口蹄疫与幼畜腹泻、生产安全高效的基因工程联合疫苗提供试验依据。[方法]通过基因工程技术,把O型口蹄疫病毒VP1双拷贝21-40(20aa)与141-160(20aa)表位肽基因——2020VP1与产肠毒素大肠杆菌LTB连接到表达载体上,在大肠杆菌中表达出具有抗O型口蹄疫病毒与产肠毒素大肠杆菌双重作用的融合蛋白。[结果]运用基因工程技术构建了融合表达载体r2020-B-2020。转化宿主菌BL21(DE3)RIL后的表达产物经SDS-PAGE分析,结果显示,重组融合蛋白的分子量约为41 kD,表达量较高。动物实验表明,融合蛋白能够诱发兔体产生较强的抗FMDV中和抗体,免疫豚鼠在低浓度FMDV刺激下产生特异性T淋巴细胞增殖反应,说明融合蛋白可以诱导机体产生有效的抗FMDV细胞及体液免疫反应。融合蛋白能够与霍乱毒素CTB抗体特异结合,免疫雌鼠能够抵抗一定强度的大肠杆菌毒株攻击。[结论]融合蛋白可以同时诱导机体产生较好的抗FMDV及ETEC免疫应答反应,具有开发成为FMDV与ETEC基因工程疫苗的应用价值。The research aimed to provide the experimental basis for preventing foot-and-mouth disease and diarrhea of young animals and producing secure and efficient gene engineering combined vaccines.By genetic engineering technology,two-copy gene of 21-40（20aa）and 141-160（20aa） epitope peptides of Type O of food-and-mouth disease virus VP1-2020VP1 was connected with enterotoxigenic E.coli LTB gene on expression vector and a fusion protein that against type O of food-and-mouth disease virus and enterotoxigenic E.coli was expressed in E.coli.A fusion expression vector r2020-B-2020 was constructed by using genetic engineering technology.SDS-PAGE analysis was conducted on the expression product after transforming host bacteria BL21（DE3） RIL and the results showed that the molecular weight of the recombinant fusion protein was about 41 kD and had higher expression quantity.Animal experiment showed that the fusion protein could induce stronger neutralization antibody against FMDV in rabbits and the multiplication response of specific T Lymphocytes in the immunized guinea pigs,which indicated that the fusion protein could induce the production of effective cells against FMDV and humoral immune response in the body.The fusion protein could be specifically combined with CTB antibody of cholera toxin and the immunized female rats could resist the attack from virus strains of E.coli to a certain extent.The fusion protein could induce better immune response against FMDV and ETEC simultaneity and had application value of developing into genetic engineering vaccine of FMDV and ETEC.

关 键 词：口蹄疫病毒 VP1 细胞表位 产肠毒素大肠杆菌 LTB 免疫应答 

分 类 号：Q503[生物学—生物化学]