摘要
将克隆到pUC119中的传染性喉气管炎病毒 (ILTV)糖蛋白gB基因 ,通过EcoRI位点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_BlueDNA)共转染Sf9昆虫细胞 ,经 3轮蚀斑纯化 ,获得重组病毒并命名为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Sf9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。
The gB gene of infectious laryngotracheitis virus(ILTV)strain WG was subcloned into the Baculovirus transfer vector pVL1393 and a recombinant vector rpVLgB was generated. Co_transfection of Sf9 insect cells with rpVLgB and Baculovirus linear DNA was performed (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.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2001年第5期321-324,共4页
Chinese Journal of Preventive Veterinary Medicine
