【作者】 张绍杰； 倪健强； 孟松树； 王柳； 仇华吉； 王云峰； 童光志；

【Author】 ZHANG Shaojie, NI Jianqiang, MENG Songshu, WANG Liu, QIU Huaji, WANG Yunfeng, and TONG Guangzhi(National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, CAAS, Harbin 150001)

【机构】 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室；

【摘要】 将克隆到pUC119中的鸡传染性喉气管炎病毒糖蛋白gB基因,通过EcoRI位点再亚克隆到杆状病毒转移载体pVL1393中,构建成重组杆状病毒转移载体rpVLgB。将rpVLgB转移载体质粒与杆状病毒DNA(Bac-N-Blue DNA)共转Sf9昆虫细胞,经三轮蚀斑纯化,获得的重组病毒命名为rpVL-ILTVgB。PCR方法鉴定证明,gB基因正确插入到杆状病毒基因组中,直接免疫荧光试验和Dot-ELISA结果均表明gB基因在重组杆状病毒感染的S19昆虫细胞中获得表达,表达的gB蛋白将作为鸡传染性喉气管炎的亚单位疫苗和诊断抗原。更多还原

【Abstract】 The gB gene of infectious laryngotracheitis virus (ILTV) strain WG was sub-cloned into the Baculovirus transfer vector pVL1393 and a recombinant vector rpVLgB was generated. Co-transfection of SF9 insect cells was performed with rpVLgB and Baculovirus linear DNA (BAC-N-Blue DNA). After three times purification, a recombinant baculovirus, designated as rpVL-ILTVgB was obtained. Expression of ILTV gB in rpVL-ILTVgB infected SF9 cells was detected by direct immunofluorescence and Dot-ELISA. The expressed glycoprotein B will be used as subunit vaccine or diagnostic antigen.更多还原