Construction and characterization of recombinant fowlpox virus co-expressing F and HN genes of newcastle disease virus and gB gene of infectious larygnotracheitis virus |
| |
Authors: | Hui-Ling SUN Yun-Feng WANG De-Yuan MIAO Pei-Jun ZHANG Hai-Dong ZHI Ling-Long XU Mei WANG Guang-Zhi TONG Ming WANG |
| |
Institution: | aCollege of Veterinary Medicine, China Agricultural University, Beijing 100094, China;bNational Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, CAAS, Harbin 150001, China;cInstitute for Animal Husbandry and Veterinary Science, Beijing Academy of Agricultural and Forestry Sciences, Beijing 100089, China |
| |
Abstract: | The Fusion (F) and Haemagglutinin-Neuraminidase (HN) genes of Newcastle disease virus (NDV) and the glycoprotein B (gB) gene of infectious laryngothracheitis virus (ILTV) as well as a LacZ reporter gene were all inserted into a nonessential gene of fowlpox virus (FPV) 017 strain by homologous recombination. The NDV and ILTV genes were each under the control of a fowlpox virus immediate early/late promoter (LP2EP2), whereas the LacZ reporter gene expression cassette was regulated by a P11 late promoter. A recombinant FPV harboring the F, HN and gB genes as well as the LacZ gene, designated as rFPV-F/HN/gB/LacZ, was obtained after ten cycles of blue plaque purification. The presence of the NDV and ILTV genes was confirmed by PCR. The expression of the recombinant proteins in rFPV-F/HN/gB/LacZ was characterized by Western blot (F and gB proteins) and indirect immunofluorescence tests (F, HN and gB proteins). The results demonstrated that all four foreign proteins, which were encoded within a 10-kb gene fragment, could be expressed authentically and efficiently. Compared with the parental virus, rFPV-F/HN/gB/LacZ showed no obvious difference with respect to virus replication and cytopathogenic effects in the cell culture of chicken embryo fibroblasts (CEF). Overall, this study suggests that FPV can be a useful live virus vector for the expression of multiforeign genes against multiple avian pathogens. |
| |
Keywords: | recombinant fowlpox virus Newcastle disease virus infectious laryngothracheitis virus F gene HN gene gB gene |
本文献已被 ScienceDirect 等数据库收录! |
| 点击此处可从《生物工程学报》浏览原始摘要信息 |
| 点击此处可从《生物工程学报》下载免费的PDF全文 |
|