【摘要】：分析獲得pm0979、ompH基因的主要抗原表位,并將2種基因以3種不同長度的疏水性linker進(jìn)行拼接融合(標(biāo)記為pm0979-L6-ompH、pm0979-L10-ompH、pm0979-L15-ompH),分別克隆于pet-30a中構(gòu)建原核表達(dá)載體,并轉(zhuǎn)化至BL21(DE3)感受態(tài)細(xì)胞進(jìn)行表達(dá)獲得融合蛋白。以融合蛋白免疫小鼠后進(jìn)行牛源多殺性巴氏桿菌(Pm)CQ2株、Pm B型攻毒試驗(yàn),分別測定小鼠體內(nèi)抗體產(chǎn)生情況以及對小鼠的交叉保護(hù)效果。結(jié)果顯示:3種融合蛋白在小鼠體內(nèi)都能誘導(dǎo)較高的抗體水平,但不同長度的linker對小鼠保護(hù)性不同,其中融合蛋白rPM0979-L10-OMPH、rPM0979-L15-OMPH對Pm CQ2株攻毒的小鼠保護(hù)率達(dá)70%,對Pm B型攻毒小鼠保護(hù)率為30%。結(jié)果表明:重組的pm0979-ompH融合基因疫苗具有良好的交叉免疫保護(hù)力;同時(shí),不同長度linker的基因疫苗對小鼠的保護(hù)率有一定影響。
[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.
【作者單位】： 西南大學(xué)動(dòng)物科技學(xué)院/重慶市牧草與草食家畜重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家現(xiàn)代農(nóng)業(yè)(肉牛牦牛)產(chǎn)業(yè)技術(shù)體系建設(shè)專項(xiàng)基金資助項(xiàng)目(CARS-38)
【分類號(hào)】：S852.61
，

本文編號(hào)：2329702