鹅细小病毒VP3基因与鹅γ干扰素基因在重组禽痘病毒中的共表达

Co-expression of GPV-VP3 gene and goose interferonγgene in recombinant fowlpox virus

为了构建共表达鹅细小病毒(GPV)VP3基因与鹅IFNγ基因的重组禽痘病毒,本研究将GPV-VP3基因和鹅IFNγ基因克隆到禽痘病毒转移载体中,构建串联基因的禽痘病毒转移载体pSY-GoIFNγ-VP3,采用脂质体法将其转染禽痘病毒感染的鸡胚成纤维细胞(CEF),经蓝斑法筛选重组病毒,并进行8轮蚀斑纯化,以及PCR鉴定和间接免疫荧光(IFA)分析。结果表明,所获得的重组病毒rFPV-GoIFNγ-VP3能稳定表达外源基因。本研究为进一步开展GPV重组禽痘病毒疫苗的研制奠定了基础。

To construct the recombinant fowlpox virus（FPV） co-expressing VP3 gene of goose par- vovirus and goose interferon γ gene, the GPV-VP3 gene and interferon γ gene were cloned into pSY-681（pSY-GoIFNγ-VP3） ,them transfected chicken embryo fibroblast（CEF） cells pre infected with FPV（S-FPV 017） by lipofectin method, pSY-GoIFNγ-VP3 was selected by the blue virus plagues,and identified by PCR and indirect immune-fluorescence test. The results showed that the two target genes were expressed in FPV infected cells. This study laid the foundation for the fur- ther development of GPV recombinant fowl pox virus vaccine.