利用PiggyBac转座系统构建稳定表达鹅RIG-I的DF-1细胞系

摘要

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.%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。