新城疫病毒F基因与传染性法氏囊病毒VP0基因在鸡痘病毒中共表达

目的 新城疫病毒（NDV）F基因和传染性法氏囊病病毒（IBDV）VP0基因在鸡痘病毒中表达,以便研制新城疫和传染性法氏囊多价重组疫苗。方法 利用鸡痘病毒282E4株非必需区TK基因构建的表达载体PU-TA2,在已存在的复合启动子p7.5ATI下游同向插入F基因,反向插入复合启动子P7.5ATI,并在其下游反向插入VP0基因,构建由2个相同的复合启动子在相反方向分别表达F和VP0蛋白的PATI2-F-VP0表达载体。使其与鸡痘病毒282E4株共转染CEF细胞后,用BUDR法筛选重组病毒,通过间接免疫荧光法检测阳性重组病毒。用Western blot法检测F和VP0蛋白。结果 在重组病毒株中,用Western blot法可检测到F和VP0蛋白。结论 已成功地获得可同时表达F蛋白和VP0蛋白的重组鸡痘病毒。

Co-expression of Newcastle Disease Virus Gene and Infections Bursal Disease Virus Vp0 Gene in Fowlpox Virus

Objective To express Newcastle disease virus (NDV) F gene and infectious bursal disease virus Vp0 gene in fowlpox virus and provide a basis for the preparation of polyvalent recombinant ND and IBD vaccine. Methods An expression vector pAT2 - F - VpO was constructed by inserting NDV F and IBDV VpO genes into the recombinant plasmid pUTA2 containing the TK gene of nonessential region of fowlpox virus was 282E4 strain and 2 p7.5ATI promoters introduced at opposite directions.The constructed plasmid pAT12 - F - Vp0 was transfected into CEF cells infected with FPV 282 E4 strain, and the recombinants were screened out by BUDR method and detected by indirect immunofiuorescence method. F and Vp0 proteins were confirmed Western blot. Results Recombinant fowlpox virus was successfully constructed, and both F and Vp0 proteins were confirmed by Western blot. Conclusion F and Vp0 proteins were expressed in recombinant fowlpox virus at the same time and recognized by corresponding PcAbs.