A 1.66 kb expected S1 gene fragment of avian infectious brochitis virus (IBV) H strain was amplified by RT PCR. Its PCR/RFLP pattern was similar to IBV D 41 strain and M 41 strain when digested using BstYI, HaeⅢ and ...A 1.66 kb expected S1 gene fragment of avian infectious brochitis virus (IBV) H strain was amplified by RT PCR. Its PCR/RFLP pattern was similar to IBV D 41 strain and M 41 strain when digested using BstYI, HaeⅢ and EcoRI respectively. IBV H strain was thought as Massachusetts serotype.展开更多
文摘猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)纤突蛋白S1能够介导宿主抵御PEDV感染的中和抗体的产生,是PEDV基因工程疫苗研究的重要靶抗原。将PEDV的S1基因克隆至杆状病毒基因组中,构建包含PEDV S1基因的重组杆状病毒,使S1蛋白表面展示表达。以该重组病毒作为活载体疫苗皮下接种BALB/c小鼠,验证S1蛋白在重组杆状病毒中的表达及其免疫原性。Western blotting分析被重组杆状病毒感染的Sf9细胞中,PEDV S1蛋白能有效表达且呈现大小分别约120 k D和150 k D的2条带,其中前者至少比预期大30 k D左右,推测重组S1蛋白在Sf9细胞中被高度糖基化修饰。重组杆状病毒的胶体金免疫电子显微镜观测结果显示S1蛋白展示于杆状病毒表面;间接ELISA试验结果证明重组杆状病毒可以诱导产生PEDV特异性抗体,抗体效价达到1∶5 000。血清中和试验及淋巴细胞增殖试验结果证明,该重组杆状病毒能够激发有效的体液免疫及细胞免疫反应,S1蛋白在重组杆状病毒表面展示表达并具有免疫原性。
文摘A 1.66 kb expected S1 gene fragment of avian infectious brochitis virus (IBV) H strain was amplified by RT PCR. Its PCR/RFLP pattern was similar to IBV D 41 strain and M 41 strain when digested using BstYI, HaeⅢ and EcoRI respectively. IBV H strain was thought as Massachusetts serotype.
