表达鸡Ⅱ型干扰素重组鸡痘病毒的构建及其特性分析  

Construction and Characterization of Recombinant Fowlpox virus Expressing Chicken Type Ⅱ Interferon

作　　者：孙永科[1] 王云峰[1] 智海东[1] 刘胜旺[1] 王玫[1] 童光志[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室

出　　处：《农业生物技术学报》2005年第1期81-85,共5页Journal of Agricultural Biotechnology

基　　金：国家高技术研究与发展计划(863)项目(No.2001AA213041)资助

摘　　要：将鸡Ⅱ型干扰素(ChIFNγ)基因插入到鸡痘病毒转移载体pSY681中,获得重组转移载体pSY681-ChIFNγ。将pSY681-ChIFNγ转染已感染亲本鸡痘病毒S-FPV-017株的鸡胚成纤维细胞,使其在鸡胚成纤维细胞内与鸡痘病毒基因组发生同源重组,产生表达ChIFNγ的重组鸡痘病毒rFPV-ChIFNγ。在含有X-gal的营养琼脂培养基上进行蓝斑筛选,通过PCR和间接免疫荧光等实验证实获得纯化的、表达ChIFNγ的重组鸡痘病毒。将在鸡胚成纤维细胞中培养72h的rFPV-ChIFNγ上清接种小鼠成纤维细胞(L929),通过细胞病变抑制实验证明重组ChIFNγ具有抑制水疱性口炎病毒复制的活性,培养上清的抗病毒效价为2048U/mL。Chicken type Ⅱ interferon(ChIFNγ ) gene was cloned into the transfer vector pSY681 of Fowlpox virus and the derived recombinant plasmid was designated as psy681-ChIFNγ . Chicken embryo fibroblasts cells pre-infected with wild type Fowlpox virus S-FPV-017 were transfected with pSY681-ChIFNγ . Recombinant Fowlpox virus rFPV-ChIFNγ expressing ChIFNγ was screened by plaque assay, PCR detection and immunofluoresce assay. The cytopathic effect (CPE) induced by vesicular stomatics virus in L929 cell line was suppressed by the cell culture supernatant of rFPV-ChIFNγ with the titer of 2 048 U per milliliter.

关 键 词：IFNΓ 表达 干扰素 重组鸡痘病毒 蓝斑 培养上清 间接免疫荧光 鸡胚成纤维细胞 转移载体 水疱性口炎病毒 

分 类 号：S858[农业科学—临床兽医学] S852[农业科学—兽医学]