马岗鹅RIG-Ⅰ基因扩增及序列分析  

Amplification and Sequence Analysis of RIG-Ⅰ Gene in Magang Goose

作　　者：叶芷羽 李伟强 刘智婷 庄海彬 车斯琪 陈治濠 欧阳婷 王蕊[1] 焦培荣[1,2,3] YE Zhiyu;LI Weiqiang;LIU Zhiting;ZHUANG Haibin;CHE Siqi;CHEN Zhihao;OUYANG Ting;WANG Rui;JIAO Peirong(College of Veterinary Medicine,South China Agricultural University,Guangzhou,Guangdong 510642;Key Laboratory of Animal Vaccine Development,Ministry of Agriculture and Rural Affairs,Guangzhou,Guangdong 510642;Key Laboratory of Zoonoses Control and Prevention of Guangdong,Guangzhou,Guangdong 510642)

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]农业农村部兽用疫苗创制重点实验室,广东广州510642 [3]广东省动物源性人兽共患病预防与控制重点实验室,广东广州510642

出　　处：《中国家禽》2021年第12期26-32,共7页China Poultry

基　　金：国家自然科学基金项目(31872497、32072844)。

摘　　要：为研究马岗鹅维甲酸诱导基因Ⅰ(Retinoic acid inducible gene-Ⅰ,RIG-Ⅰ)的基因序列特征,试验扩增了马岗鹅RIG-Ⅰ基因,分析了马岗鹅RIG-Ⅰ基因序列并表达了其蛋白。结果显示:马岗鹅RIG-Ⅰ基因cDNA的开放阅读框全长为2 805 bp,编码934个氨基酸,蛋白分子量约为106.63 ku;马岗鹅RIG-Ⅰ蛋白具有2个串联的N端半胱天冬酶募集结构域、DExD/H-box RNA解旋酶结构域和C端结构域;马岗鹅RIG-Ⅰ与其它鸟类亲缘关系较近,与哺乳动物亲缘关系次之,与鱼类亲缘关系较远;马岗鹅RIG-Ⅰ蛋白结构以α-螺旋为主(占比50.21%),无规则卷曲次之(占比37.69%),延伸链较少(占比12.10%);马岗鹅RIG-Ⅰ真核表达质粒成功表达了约106 ku的蛋白。研究成功鉴定了马岗鹅RIG-Ⅰ基因并表达其蛋白,为进一步制备该蛋白的特异性抗体和研究鹅RIG-Ⅰ在抗病毒天然免疫中的功能奠定了基础。To study the sequence characteristic of retinoic acid inducible gene-Ⅰ(RIG-Ⅰ) in Magang goose,the full-length RIG-Ⅰ was amplified,sequenced and then expressed in 293 T cells.The results showed that the full-length open reading frame of RIG-Ⅰ gene was 2 805 bp encoding 934 amino acids,and the molecular weight of the coding protein was about 106.63 ku;RIG-Ⅰmainly included 2 of caspase activation and recruitment domain(CARD),DExD/H-box helicase domain and C-terminal domain(CTD);RIG-Ⅰ of Magang goose was closely related to RIG-Ⅰ gene of the other birds,next with mammals and the farthest with fish;RIG-Ⅰ protein consisted of α-helix(50.21%),irregular coil(37.69%) and extended strand(12.10%);The eukaryotic expression plasmids of RIG-Ⅰ was successfully expressed a 106 ku protein in 293 T cells.It′s suggested that the identified RIG-Ⅰof Magang goose and its protein would lay the foundation for the further investigation of the function of goose RIG-I in innate immune and preparation of specific antibodies.

关 键 词：马岗鹅 RIG-Ⅰ基因 基因扩增 序列分析 蛋白表达 

分 类 号：S855.3[农业科学—临床兽医学]