共表达口蹄疫病毒vp1基因和山羊IFN-γ基因的重组山羊痘病毒的构建

Construction of recombinant capripox virus co-expressing FMDV vp1 and goat IFN-γ

将山羊IFN-γ基因插入到含有口蹄疫病毒(FMDV)vp1基因的山羊痘病毒(GPV)转移载体PTK-vp1中,获得含有FMDVvp1基因和山羊IFN-γ基因的重组转移载体pTK-vp-IFNγ,以本实验室构建的表达β-半乳糖苷酶基因(LacZ)的山羊痘重组病毒rAV41-LacZ为亲本毒,与转移载体pTK-vp-IFNγ共同转染犊牛睾丸细胞进行同源重组,通过7轮反向蓝白斑筛选,产生重组山羊痘病毒rAV41-VP-IFNγ,通过PCR和间接免疫荧光实验证实获得了能够稳定表达FMDVvp1和山羊IFN-γ的重组山羊痘病毒。

We constructed the recombinant transfer vector pTK-VP-IFNγ by inserting the goat IFN-γ gene into pTK-VP1 which contains the FMDV viol gene, The pTK-VP-IFN γ vector was purified and transfected by lipofection into bovine testicle cells that was infected with rAV41-LacZ expressing Escherichia coli β -Galactosidase （LacZ）. A recombinant virus co-expressing the goat IFN-γ gene and FMDV viol gene was generated after seven cycles of white plaque purification and PCR identification. Expression and immunogenicity of the viol and IFN- γ were analyzed by indirect immunofluorescence test.