摘要
为了构建枯草芽孢杆菌芽孢表面展示猪轮状病毒(PoRV)抗原蛋白VP8的重组菌株,本试验将包含猪轮状病毒VP8基因和枯草芽孢杆菌芽孢外衣壳蛋白基因cot C同时插入枯草芽孢杆菌穿梭质粒pDG364,融合基因以同源双交叉重组的方式整合入枯草芽孢杆菌基因组,构建了芽孢表面展示猪轮状病毒VP8蛋白的重组菌枯草芽孢杆菌CV,利用淀粉酶活性分析、PCR鉴定及免疫荧光显微镜观察对重组菌进行鉴定。结果表明,融合基因成功整合入受体菌基因组,在重组菌形成芽孢时融合基因得到正确表达,免疫荧光显微镜观察重组芽孢可见特异性荧光信号,在芽孢表面展示的VP8蛋白具有反应原性。
To constructed a recombination strain with spore surface display heterologous antigen protein VP8 of porcine rotavirus.A shuttle vector was constructed for the spore surface display of the spike-protein VP8 from porcine rotavirus,using spore coat protein cotC as a carrier.The fusion gene cotC-VP8 was integrated into B.subtilis 168 chormosome at the amyE locus by a double crossover enent.Constructed a recombination strain B.subtilis CV with spore surface display heterologous antigen protein VP8 of porcine rotavirus.B.subtilis CV was tested by amylase activity assay,PCR,and the spore was observed under the fluorescence immunossay.The results indicated that the fusion gene was inserted in amyE and VP8 was successed displayed on the spore with biological activity.
作者
冯杰
潘康成
FENG Jie;PAN Kangcheng(Fushun Animal Husbandry and Veterinary Technology Extension Center,Zigong 643200,China;Fushun Animal Disease Control Center,Zigong 643200,China;College of Veterinary Medicine,Sichuan Agricultural University,Chengdu 611130,China)
出处
《中国兽医卫生》
2025年第4期23-27,31,共6页
China Veterinary Hygiene
