传染性喉气管炎病毒gB基因和新城疫病毒F基因在重组禽痘病毒中共表达

在重组禽痘病毒中表达多个禽类病原的主要免疫原基因是构建多价基因工程疫苗的前提，但相关研究很少。在表达传染性喉气管炎病毒(ILTV)gB基因重组禽痘病毒的转移载体的基础上，构建了含有ILTV gB基因和新城疫病毒(NDV)F基因的重组禽痘病毒转移载体pSY-gB-F，采用脂质体转染禽痘病毒感染的鸡胚成纤维(CEF)细胞后，通过蓝斑试验筛选出重组禽痘病毒(rFPv-gB-F)，并进行了6轮蚀斑纯化。Western-blot试验和间接免疫荧光试验证明ILTV gB基因和NBVF基因在rFPV-gB-F感染的CEF细胞中获得表达。为传染性喉气管炎、新城疫与鸡痘活载体多价疫苗的研制奠定基础。

Co-expression of Glycoprotein B (gB) of Infectious Laryngotracheitis Virus and Fusion Protein of Newcastle Disease Virus in a Recombinant Fowlpox Virus

Fowlpox virus has been used as vector to successfully express protective immunogen genes of many avian virus, while co-expression immunogens from more virus was seldom reported. Based on the construction of a recombinant Fowlpox Virus expressing gB gene of Infectious Laryngotracheitis Virus (ILTV) previously, a transfer vector containing F gene of Newcastle Disease Virus (NDV) and gB of ILTV was constructed, after transfection chicken embryo fibroblast (CEF) cells infected with FPV (S-FPV-017) by lipofectin method, recombinant FPV (designated as rFPV-gB-F) was screened by identifying blue plagues. gB and F gene inserted in the genome was identified by PCR, and expression of gB gene and F gene was detected by Western blotting and indirect immuno-fluorescence test. The results lay foundation for the development of tri-valent vaccine against ILT, ND and FP, and the immune potency of the recombinant virus needs further evaluation.