J亚群禽白血病毒gp85基因表达及初步应用  

EXPRESSION AND APPLICATION OF AVIAN LEUKOSIS VIRUS SUBGROUP J GP85

作　　者：张伟伟[1,2] 杨宗伟[1] 陈宗艳[1] 王超[1] 李传峰[1] 刘光清[1] 

机构地区：[1]中国农业科学院上海兽医研究所,上海200241 [2]浙江师范大学化学与生命科学学院,金华321004

出　　处：《中国动物传染病学报》2011年第1期17-21,共5页Chinese Journal of Animal Infectious Diseases

基　　金：863计划(2011AA10A208);公益性行业(农业)科研专项资助(201003012)

摘　　要：J亚群禽白血病毒(Avian leukosis virus subgroup J,ALV-J)的囊膜糖蛋白GP85是亚群特异性抗原。将ALV-J的gp85基因克隆至原核表达载体pET-30a(+)EcoRⅠ和SalⅠ两个酶切位点之间,经IPTG诱导在大肠杆菌中以His-Tag融合蛋白的形式获得了高效表达,Western blot显示该表达产物具有生物学活性。GP85蛋白经纯化、复性后免疫家兔,制备了抗ALV-J GP85的多克隆抗体。以纯化的蛋白为抗原包被ELISA检测板,对疑似血清样本进行了实验室检测,检出率较高。本研究为ALV-J的诊断和流行病学调查奠定了基础。The surface subunit GP85 is encoded by env gene,which determines the subgroup of Avian leukosis virus subgroup J（ALV-J）.The gp85 gene of ALV-J was amplified by PCR technique and cloned into pET-30a vector at sites of EcoR I and Sal I.The recombinant plasmid was transformed into E.coli BL21（DE3） and induced with IPTG.The result showed that the gp85 gene had been well expressed with His-Tag.The expressed product was analyzed by Western blot.Rabbits were immunized with the purified and renatured GP85 fusion protein for preparing the anti-ALV-J GP85 antibody.An ELISA method was developed based on the recombinant GP85 protein to detect sera of ALV-J infected chickens and may be used in further for diagnosis of ALV-J infection and epidemiology investigation.

关 键 词：J亚群禽白血病毒 囊膜糖蛋白 ELISA 

分 类 号：S852.659.3[农业科学—基础兽医学] Q786[农业科学—兽医学]