表达鸡传染性法氏囊病病毒VP_2重组鸡痘病毒的构建  

Recombination of Fowlpox Virus to Express the VP_2 Antigen of Chicken Infectious Bursal Disease Virus

作　　者：刘小军[1] 陈楠[1] 陈立栋[1] 周胜花[1] 陈永福[1] 

机构地区：[1]中国农业大学农业生物技术国家重点实验室

出　　处：《农业生物技术学报》2000年第2期186-189,共4页Journal of Agricultural Biotechnology

摘　　要：含IBDV保护性抗原VP2的质粒，以SphⅠ消化，然后用T4噬菌体DNA聚合酶将末端补平，产物纯化后再以SalⅠ消化一次，经再次纯化后与SmlⅠ和SalⅠ分步酶切的鸡瘟病毒插入载体pFG1175－1相连，使VP2基因顺向插入到栽体P75启动子下游，得到合IBDVVP2基因的重组插入载体pFGVP2。将该质粒用脂质体介导转染，用禽痘病毒疫苗株FPV282E4感染3～4h的鸡胚成纤维细胞，经多次蓝斑纯化后，得到稳定的重组病毒，间接免疫荧光法证明，重组病毒在细胞中表达了IBDVVP2。The combinant plasmid pEGMVP2 that contains the gene of protective antigen VP2 of IBDV was digested with SphI.The 5' end was flushed by T4 DNA polymerase. The purified product was digested with Sall again. The segment with 1 580 bp in length was recovered and inserted into the downstream of P7. 5 promotor of insertion vector pEG1175-1 digested with Smal and Sa/I previously.The recombinant vector was designated pFGVP2. The CEF cell cultures which had infected with FPV282E4 for 3-4 hours were transfected with pFGVP2 plasmid DNA. Recombinant PFV with blue plaques were selected and purified 3 times in cultures overlaid with agarose containing X-gal. The expression of IBDV VP, in recombinant FPV was confirmed by immunofluorsence assay with antibody produced against IBDV.

关 键 词：鸡传染性法氏囊病病毒 VP2基因 重组鸡痘病毒 

分 类 号：S858.312.6[农业科学—临床兽医学]