表达鹅γ干扰素重组禽痘病毒的构建及其抗病毒活性的初步研究  

Expression of goose interferon-γ gene from recombinant fowlpox virus and preliminary study on its inhibitory activity against goose parvovirus

作　　者：张紫璇 韩明子 高旭[1] 李远超 孟媛 李卓昕 王美淇 刘晋宇[1] ZHANG Zi-xuan;HAN Ming-zi;GAO Xu;LI Yuan-chao;MENG Yuan;LI Zhuo-xin;WANG Mei-qi;LIU Jin-yu(Department of Veterinary Medicine,Agricultural College,Yanbian University,Yanji 133000,China;Yanbian Prefecture Animal Disease Prevention and Control Center,Yanji 133000,China)

机构地区：[1]延边大学农学院动物医学系,吉林延吉133000 [2]延边州动物疫病预防控制中心,吉林延吉133000

出　　处：《中国预防兽医学报》2018年第12期1163-1167,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金项目(31460661)

摘　　要：为研究表达鹅γ干扰素(GoIFN-γ)基因重组禽痘病毒的抗病毒活性,本研究构建了在禽痘病毒(FPV)早晚期启动子LP2EP2控制下的IFN-γ基因重组禽痘病毒转移载体,将其转染至亲本禽痘病毒S-FPV-017预感染的鸡胚成纤维细胞(Chicken embryo fibroblasts,CEF),使其与禽痘病毒基因组进行同源重组,经过9轮筛选和纯化并进行PCR和间接免疫荧光检测,获得能稳定表达外源基因的重组病毒r FPV-IFNγ-LacZ。利用细胞病变抑制法检测抗病毒活性结果显示,在鸡胚成纤维细胞中重组禽痘病毒表达的IFN-γ对鹅细小病毒的复制具有显著抑制作用。本研究为制备共表达保护性抗原和γ干扰素基因的重组基因工程疫苗研究奠定基础。In order to study the antiviral activity of recombinant fowlpox virus (FPV) expressing goose interferon-γ (GoIFN-γ), a recombinant fowlpox virus transfer vector containing GoIFN-γ gene under the control of the early and late promoter of the FPV LP2EP2 was constructed in this study. The transfer vector was transfected into the chicken embryo fibroblasts (CEF) pre-infected with FPV S-FPV-017 for homologous recombination. After several rounds of blue plaque screening and purification, a recombinant FPV stably expressing GoIFNγ and LacZ genes (rFPV-IFNγ-LacZ) was obtained and identified by PCR and indirect immunofluorescence assay. The results of cytopathic inhibition assay on the antiviral activity of the recombinant virus showed that the GoIFNγ expressed from rFPV-IFNγ-LacZ infected CEF was able to inhibit the replication of goose parvovirus. This study laid the foundation for the preparation of recombinant FPV vaccine co-expressing of protective antigen and gamma interferon.

关 键 词：Γ干扰素 重组禽痘病毒 鹅细小病毒 

分 类 号：S852.65[农业科学—基础兽医学]