本文選題：豬O型口蹄疫病毒　切入點(diǎn)：表位　出處：《甘肅農(nóng)業(yè)大學(xué)學(xué)報(bào)》2017年01期

【摘要】：【目的】制備豬O型FMDV廣譜多表位疫苗,并篩選最適合的免疫佐劑.【方法】選取O型FMDV 3個(gè)毒株VP1蛋白的優(yōu)勢(shì)表位,設(shè)計(jì)并合成重復(fù)串聯(lián)表位基因3FoEN2,并克隆豬IgG重鏈恒定區(qū)基因.利用BamHⅠ,EcoRⅠ等位點(diǎn)將2個(gè)基因依次克隆到pProEX-HTb載體,構(gòu)建重組質(zhì)粒pE-IgG并轉(zhuǎn)化大腸桿菌BL21(DE3)感受態(tài)細(xì)胞.以IPTG誘導(dǎo)表達(dá)得到融合蛋白pE-IgG,經(jīng)SDS-PAGE電泳分析,Western-blotting鑒定.分別用5種佐劑ISA206、ISA201、IMS1313、603、ISA61乳化融合蛋白配制疫苗免疫BALB/c雌鼠,間接ELISA方法測(cè)定抗體水平.【結(jié)果】重組蛋白以包涵體形式正確表達(dá),大小為45kU,且能與感染O型FMDV的豬的陽(yáng)性血清發(fā)生特異性免疫反應(yīng);ISA201佐劑試驗(yàn)組刺激機(jī)體產(chǎn)生的抗體水平最高.【結(jié)論】pE-IgG蛋白具有很強(qiáng)的免疫原性,與ISA201佐劑混合制備成的豬O型口蹄疫病毒多表位疫苗可刺激動(dòng)物機(jī)體產(chǎn)生高水平抗體,是具有良好開(kāi)發(fā)前景的疫苗.
[Abstract]:[objective] to prepare porcine type O FMDV broad-spectrum multiepitope vaccine and to screen the most suitable immune adjuvant. [methods] the dominant epitopes of VP1 protein of three strains of FMDV were selected.The repeat tandem epitope gene 3FoEN2 was designed and synthesized and cloned into porcine IgG heavy chain constant region gene.Two genes were cloned into pProEX-HTb vector by allele of BamH 鈪，

本文編號(hào)：1711296