表达鸡白细胞介素18重组禽痘病毒的构建及其表达产物生物学活性的检测  

Construction of Recombinant Fowlpox Virus Expressing Chicken Interleukin-18 and Its Biologic Activity Detection

作　　者：陈红英[1] 黄青云[2] 崔保安[1] 李新生[1] 管倩[1] 刘金朋[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002 [2]华南农业大学兽医学院,广东广州510642

出　　处：《华北农学报》2008年第6期6-10,共5页Acta Agriculturae Boreali-Sinica

基　　金：国家“十一五”科技支撑计划专项(2006BAD06A08)

摘　　要：为构建表达鸡IL-18的重组禽痘病毒,将含痘病毒启动子LP2EP2驱动的鸡IL-18基因插入到禽痘病毒转移载体pSY681中,获得重组转移载体pSY681/ChIL-18。将pSY681/ChIL-18转染已感染亲本禽痘病毒S-FPV-017株的鸡胚成纤维细胞,使其在鸡胚成纤维细胞内与禽痘病毒基因组发生同源重组,产生表达鸡IL-18的重组禽痘病毒rFPV-ChIL-18。在含有X-gal的营养琼脂培养基上进行蓝斑筛选后,对重组病毒rFPV-ChIL-18又进行了多次蚀斑克隆。以重组禽痘病毒DNA为模板,利用鸡IL-18基因特异引物进行PCR,扩增出1条约0.6 kb的带。收集含鸡IL-18蛋白的细胞上清液进行MTT试验,表达的产物能明显提高鸡淋巴细胞的转化率。In order to construct the recombinant fowlpox viruses（rFPV） expressing chicken IL-18, the chicken IL-18 cDNA was cloned into EcoR Ⅰ site of pSY538 plasmid and then subcloned into Sfi Ⅰ site of pSY681 plasmid containing LacZ gene to generate the recombinant plasmid pSY681/ChIL-18. The recombinant plasmid was transfected on the chicken embryo fibroblasts cell（ CEF）that was pre-infected with S-FPV-017 .By screening of blue plaques on the CEF overlaid with agar containing X-gal,the rFPV-ChIL-18 recombinants were obtained and identified by PCR. Recombinant chicken IL-18 induced in vitro the proliferation of ConA-stimulated chicken splenocytes in MTT assay.

关 键 词：鸡IL-18基因 重组禽痘病毒 构建 生物学活性 

分 类 号：S852.4[农业科学—基础兽医学] Q786[农业科学—兽医学]