小鼠LRRFIP1基因的克隆及LRRFIP1天然免疫模式识别功能的研究  

Cloning of mouse LRRFIP1 gene and study on functions of LRRFIP1 in innate immunity pattern recognition

作　　者：李婷婷[1] 何小兵[1] 贾怀杰[1] 陈国华[1] 曾爽[1] 房永祥[1] 金启旺 景志忠[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部兽医公共卫生重点实验室,甘肃兰州730046

出　　处：《中国兽医科学》2015年第6期649-653,共5页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31302072;31372423;30871884)

摘　　要：应用RT-PCR技术,从小鼠脾淋巴细胞中成功克隆了LRRFIP1基因,并对其遗传演化关系及LRRFIP1的分子结构特征进行了分析。结果显示,克隆的基因开放阅读框全长2 190bp,编码729个氨基酸,与大鼠、猪、牛等哺乳动物LRRFIP1相应序列的同源性为90.5%~46.5%。将克隆的小鼠LRRFIP1基因亚克隆至真核表达载体pCMV-Tag 2B,转染HEK293T细胞后,双荧光素酶报告系统证实,在poly(dA-dT)和poly(dG-dC)刺激下,过表达小鼠LRRFIP1能够通过激活IRF3而显著增强IFN-β的表达。结果表明,小鼠LRRFIP1能够识别富含A/T或C/G的DNA序列,在宿主抗DNA病毒感染的天然免疫反应中可能发挥重要作用。The coding sequence of mouse LRRFIP1 gene was amplified from splenic mononuclear cells by RT-PCR.In addition,the genetic evolution and molecular structure of the mouse LRRFIP1 gene were analyzed.In result,the ORF of this sequence is 2 190 bp in size,encoding 729 amino acids.The mouse LRRFIP1 gene exhibited identity with the corresponding sequences of rat,pig,cattle,ranging from 46.5% to90.5%.The cloned LRRFIP1 gene was subcloned into the eukaryotic expression vector pCMV-Tag 2B.The recombinant plasmid was then transfected into the HEK293 Tcells.The IRF3 and IFN-βluciferase reporter systems verified that overexpression of mouse LRRFIP1 significantly induced the activity of IFN-βby activating transcription factor IRF3 with the stimulation of poly(dA-dT)and poly(dG-dC).These results indicated that mouse LRRFIP1 was able to recognize A/T or G/C-rich DNA sequence,and may play a vital role in host innate immune responses against DNA virus infections.

关 键 词：小鼠 LRRFIP1基因 结构分析 DNA识别受体 信号转导 

分 类 号：S852.4[农业科学—基础兽医学]