牛源多杀性巴氏杆菌pm0979-ompH融合表达及免疫效果
发布时间：2018-11-13 16:35
【摘要】：分析获得pm0979、ompH基因的主要抗原表位,并将2种基因以3种不同长度的疏水性linker进行拼接融合(标记为pm0979-L6-ompH、pm0979-L10-ompH、pm0979-L15-ompH),分别克隆于pet-30a中构建原核表达载体,并转化至BL21(DE3)感受态细胞进行表达获得融合蛋白。以融合蛋白免疫小鼠后进行牛源多杀性巴氏杆菌(Pm)CQ2株、Pm B型攻毒试验,分别测定小鼠体内抗体产生情况以及对小鼠的交叉保护效果。结果显示:3种融合蛋白在小鼠体内都能诱导较高的抗体水平,但不同长度的linker对小鼠保护性不同,其中融合蛋白rPM0979-L10-OMPH、rPM0979-L15-OMPH对Pm CQ2株攻毒的小鼠保护率达70%,对Pm B型攻毒小鼠保护率为30%。结果表明:重组的pm0979-ompH融合基因疫苗具有良好的交叉免疫保护力;同时,不同长度linker的基因疫苗对小鼠的保护率有一定影响。
[Abstract]:The main antigen epitopes of pm0979,ompH gene were obtained, and the two genes were spliced and fused with three hydrophobic linker of different lengths (labeled pm0979-L6-ompH,pm0979-L10-ompH,pm0979-L15-ompH). The prokaryotic expression vector was cloned into pet-30a and transformed into BL21 (DE3) competent cells to express the fusion protein. After immunizing mice with fusion protein, the, Pm B type attack test of Pasteurella multocida strain (Pm) CQ2 was carried out, and the antibody production in mice and the cross protective effect on mice were determined. The results showed that all of the three fusion proteins could induce high antibody level in mice, but different lengths of linker could protect mice. The protective rate of fusion protein rPM0979-L10-OMPH,rPM0979-L15-OMPH against Pm CQ2 strain was 70%. The protective rate against Pm B attack mice was 30%. The results showed that the recombinant pm0979-ompH fusion gene vaccine had good cross-immune protection, and the different length of linker gene vaccine had a certain effect on the protective rate of mice.
【作者单位】： 西南大学动物科技学院/重庆市牧草与草食家畜重点实验室;
【基金】：国家现代农业(肉牛牦牛)产业技术体系建设专项基金资助项目(CARS-38)
【分类号】：S852.61
，
本文编号：2329702
[Abstract]:The main antigen epitopes of pm0979,ompH gene were obtained, and the two genes were spliced and fused with three hydrophobic linker of different lengths (labeled pm0979-L6-ompH,pm0979-L10-ompH,pm0979-L15-ompH). The prokaryotic expression vector was cloned into pet-30a and transformed into BL21 (DE3) competent cells to express the fusion protein. After immunizing mice with fusion protein, the, Pm B type attack test of Pasteurella multocida strain (Pm) CQ2 was carried out, and the antibody production in mice and the cross protective effect on mice were determined. The results showed that all of the three fusion proteins could induce high antibody level in mice, but different lengths of linker could protect mice. The protective rate of fusion protein rPM0979-L10-OMPH,rPM0979-L15-OMPH against Pm CQ2 strain was 70%. The protective rate against Pm B attack mice was 30%. The results showed that the recombinant pm0979-ompH fusion gene vaccine had good cross-immune protection, and the different length of linker gene vaccine had a certain effect on the protective rate of mice.
【作者单位】： 西南大学动物科技学院/重庆市牧草与草食家畜重点实验室;
【基金】：国家现代农业(肉牛牦牛)产业技术体系建设专项基金资助项目(CARS-38)
【分类号】：S852.61
，
本文编号：2329702
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