J亚群禽白血病病毒gp85基因在重组杆状病毒中的表达及鉴定  

Expression and identification of gp85 gene of J subgroup of avian leucosis virus in baculovirus system

作　　者：倪伟[1,2] 秦立廷[1] 孙美玉[1] 高玉龙[1] 潘伟[1] 王笑梅[1] 刘思当[2] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所禽传染病研究室,兽医生物技术国家重点实验室,黑龙江哈尔滨150001 [2]山东农业大学动物科技学院,山东泰安271018

出　　处：《畜牧与兽医》2011年第8期4-7,共4页Animal Husbandry & Veterinary Medicine

基　　金：现代农业产业技术体系建设专项资金资助(nycytx-42-G3-01);哈尔滨市科技攻关计划项目(2010AA6AN034)

摘　　要：为获得J亚群禽白血病病毒(ALV-J)的gp85蛋白,将ALV-J的gp85基因克隆至pFastBac-HTA供体质粒,将其转入DH10BacTM大肠杆菌感受态细胞,使gp85基因整合到Bacmid穿梭载体中,构建重组穿梭载体Bacmid-gp85。通过脂质体介导,将重组穿梭载体Bacmid-gp85转染Sf9昆虫细胞,获得重组杆状病毒rBacgp85。Western-blot和间接免疫荧光试验(IFA)鉴定结果表明ALV-J gp85蛋白在Sf9昆虫细胞中得到正确表达,表达的重组gp85蛋白分子量约为38 ku。ALV-J gp85重组蛋白在Sf9细胞中的正确表达为其功能研究和应用提供了良好的基础。In order to obtain the gp85 protein of J subgroup of avian leucosis virus （ALV-J）, gp85 gene was amplified and cloned into pFastBae-HTA to construct the recombinant donor plasmid pFastBae-HTA-gp85. Then, the pFastBac-HTA-gp85 was transformed into DH10Bac Escherichia coli competent cells to get the recombinant shuttle plasmid Bacmid-gp85. The recombinant baculovirus Bacmid-gp85 was obtained by transfecting rBac-gp85 with Cellfectina Reagen into St9 cells. The Western-blot analysis and indirect immunofluorescence assay revealed that the recombinant protein with the molecular weight of 38 kDa was expressed in the Sf9 cells. This study provides a good basis for functional analysis of gp85 protein of ALV-J.

关 键 词：J亚群禽白血病病毒 GP85基因 杆状病毒 

分 类 号：S852.65[农业科学—基础兽医学]