利用PiggyBac转座系统构建稳定表达鹅RIG-I的DF-1细胞系  

CONSTRUCTION OF CHICKEN EMBRYO FIBROBLAST CELL LINE DF-1 STABLY EXPRESSING GOOSE RIG-I THROUGH PIGGYBAC TRANSPOSON SYSTEM

作　　者：胡跃[1] 孙英杰[2] 王晓旭[3] 仇旭升[2] 宋翠萍[2] 谭磊[2] 胡桂学[1] 丁铲[2] 

机构地区：[1]吉林农业大学动物科学技术学院,长春130118 [2]中国农业科学院上海兽医研究所,上海200241 [3]安徽农业大学动物科技学院,合肥223006

出　　处：《中国动物传染病学报》2014年第3期53-60,共8页Chinese Journal of Animal Infectious Diseases

基　　金：863计划(2011AA10A209);国家自然科学基金项目(31101822);中央级公益性科研院所基本科研业务费(2014JB08)

摘　　要：维甲酸诱导基因-I(retinoic acid inducible gene-I,RIG-I)是最近几年发现的一种细胞内模式识别受体,能够特异性的识别并结合病毒RNA,进而发挥抗病毒效应。RIG-I在进化中相对保守,但已有报道和我们的前期研究显示:家禽中鸭和鹅体内均有RIG-I受体,但鸡却缺乏RIG-I受体。为了研究这种天然缺失对鸡是否有不利影响,本研究利用PiggyBac转座系统将鹅源RIG-I转染入鸡胚成纤维细胞系DF-1,两次亚克隆后获得单个克隆,以荧光、RT-PCR及Western blot进行鉴定,最终获得稳定表达鹅RIG-I的DF-1细胞系,为进一步研究鸡的先天性免疫机制和鹅RIG-I的功能打下基础。Retinoic acid inducible gene-I （RIG-I） is a family member of cytoplasmic pattern recognition receptors （PRRs）, which acts as a sensor for RNA virus and plays an important role in antiviral innate immunity. RIG-I is evolutionally conserved among most species. However, several reports and our previous studies have identified that receptors for RIG-I exist in both ducks and geese, but not in chickens. To investigate whether this natural loss of RIG-I plays a negative role in chicken, goose RIG-I was transfected into chicken embryo fibroblast cell line DF-1 through PiggyBac transposon system. Monoclonal cells were obtained from twice sub-cloned cultures. DF-1 cells stably expressing goose RIG-I were identified by detection of fluorescence, RT-PCR and Western blot. The present study has laid the foundation for future research on mechanism of chicken innate immunity and function of goose RIG-I.

关 键 词：鹅维甲酸诱导基因-I 鸡胚成纤维细胞系DF-1 PiggyBac转座系统 

分 类 号：S852.42[农业科学—基础兽医学]