禽白血病J亚型病毒分离鉴定及其gp85基因克隆表达  

Isolation and identification of J-Avian Leukosis virus and gp85 gene cloning and expression

作　　者：李广兴[1] 杨婷[1] 潘龙[1] 任晓峰[1] 王秀荣[2] 

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030 [2]中国农业科学院哈尔滨兽医研究所,哈尔滨150001

出　　处：《东北农业大学学报》2013年第6期38-43,F0002,共7页Journal of Northeast Agricultural University

基　　金：国家自然科学基金项目(31172295;31272569);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开放基金项目(SKLVBF201207)

摘　　要：禽白血病(Avian leukosis)是由禽白血病病毒(Avian leukosis virus,ALV)和禽肉瘤病毒(Avain sacromaviurs,ASV)群中病毒引起禽类多种肿瘤性疾病统称。文章从临床疑似病鸡中经临床症状观察、病理学检查和分子生物学鉴定等方法分离出一株ALV J亚型病毒。从感染鸡病料提取前病毒DNA,根据NCBI发表ALV基因序列设计引物,扩增该病毒gp85基因,扩增出924 bp目的条带,利用pET-30a载体构建原核表达质粒,鉴定正确后转化入Rosetta中进行诱导表达,表达40 ku蛋白以包涵体形式存在,有良好免疫原性,为ALV进一步研究提供实验材料。Avian Leukosis is caused by groups of avian leukemia virus （ALV） and avian sarcoma virus （ASV）, which may lead to a variety of neoplastic diseases of fowls collectively. In this study, the ALV J subgroup virus was isolated and characterized regarding of clinic symptoms, pathological lesions and molecular biology techniques. Then, the proviral DNA of ALV-J virus was extracted from the diseased chicken materials. According to ALV-J virus gene sequences published in GenBank, specific primers were designed to clone the gp85 gene. The results showed that of the gp85 gene of 924 bp was amplified successfully.This gene was inserted into pET-30a vector resulting in a prokaryotic expression plasma pET-30a-gp85. After transformation of pET-30a-gp85 into E. coil Rosseta and induced expression, the molecular weight of recombinant gp85 protein 40 ku was expressed in the form of inclusion body. Western-blot indicated that the expressed protein has good immunogenicity.

关 键 词：禽白血病 禽白血病J亚群病毒 分离鉴定 gp85原核表达 

分 类 号：S767.5[农业科学—森林保护学] X172[农业科学—林学]