合作猪TRIF基因CDS区克隆鉴定及组织表达分析  

Cloning,Identification and Tissue Expression Analysis of TRIF Gene CDS Region in Hezuo Pigs

作　　者：胡慧慧 付盼盼 李杰[1] 闫尊强 高小莉[1] 杨姣姣[1] 黄晓宇[1,2] HU Huihui;FU Panpan;LI Jie;YAN Zunqiang;GAO Xiaoli;YANG Jiaojiao;HUANG Xiaoyu(College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;Gansu Research Center for Swine Production Engineering and Technology,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物科学技术学院,兰州730070 [2]甘肃省现代养猪工程技术研究中心,兰州730070

出　　处：《中国畜牧兽医》2025年第4期1478-1487,共10页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金资助项目(32360819);甘肃地方猪种质资源保护及品质提升(GSLK-2021-13);甘肃农业大学畜牧学养猪产业技术创新团队(GAU-XKTD-2022-25)。

摘　　要：【目的】β-干扰素TIR结构域衔接蛋白(TIR domain-containing adaptor protein inducing interferon β,TRIF)基因在机体抗病免疫中具有重要作用,本研究旨在探究合作猪TRIF基因CDS区序列特征及组织表达特点。【方法】以甘肃合作猪为研究对象,克隆TRIF基因CDS区全长序列,并对其进行生物信息学分析,采用实时荧光定量PCR技术检测合作猪TRIF基因在肺脏、肾脏、脾脏、背最长肌、盲肠、回肠、直肠、结肠共8种组织中的表达量。【结果】合作猪TRIF基因CDS区全长2 142 bp,编码713个氨基酸,与NCBI数据库中收录的野猪TRIF基因序列(登录号:NM_001315738.2)相似性为99.81%。系统进化树结果显示,合作猪与野猪亲缘关系最近,与斑马鱼亲缘关系最远。合作猪TRIF蛋白具有亲水性,无跨膜区域,不存在信号肽,且不存在N-糖基化修饰位点,其主要定位于细胞核内。TRIF蛋白的二级结构主要以无规则卷曲为主,三级结构与二级结构预测结果基本一致,该蛋白存在6个低复杂度结构域和1个卷绕线圈区域。实时荧光定量PCR检测结果表明,合作猪TRIF基因在肺脏、肾脏、脾脏、背最长肌、盲肠、回肠、直肠、结肠中均有表达,其中在背最长肌中表达量最高。【结论】本试验成功克隆出合作猪TRIF基因CDS区序列,其在背最长肌中的表达量最高,该研究结果可为后续研究合作猪TRIF基因提供参考依据。【Objective】TIR domain-containing adaptor protein inducing interferonβ(TRIF)gene played the important roles in host resistance and immunity.This study was aimed to study the sequence characteristics and tissue expression of CDS region of TRIF gene in Hezuo pigs.【Method】Gansu Hezuo pigs was taken as the research object,the full-length CDS region sequence of TRIF gene was cloned and subjected to bioinformatics analysis.The expression levels of TRIF gene were detected from 8 tissues including lung,kidney,spleen,longissimus dorsi muscle,cecum,ileum,rectum and colon through Real-time quantitative PCR.【Result】The full length of TRIF gene CDS region was 2142 bp and encoded 713 amino acids,which was 99.81%similarity to TRIF gene sequence of Sus scrofa in NCBI database(accession No.:NM_001315738.2).Phylogenetic tree results showed that Hezuo pigs were closest to Sus scrofa and furthest to Danio rerio.TRIF protein of Hezuo pigs had hydrophilicity,no transmembrane region,no signal peptide and no N-glycosylation modification sites,which was mainly located in the nucleus.TRIF protein was mainly composed by random coils of secondary structure.The tertiary structure prediction was basically consistent with the secondary structure.The protein had six low-complexity domains and a winding coil region.Real-time quantitative PCR results showed that TRIF gene of Hezuo pigs was expressed in eight tissues including lung,kidney,spleen,longissimus dorsi muscle,cecum,ileum,rectum and colon,with the highest expression in longissimus dorsi muscle.【Conclusion】This study had successfully cloned the CDS region sequence of TRIF gene in Hezuo pigs.The highest expression was found in longissimus dorsi muscle.The results could provide a valuable reference for the further study of TRIF gene in Hezuo pigs.

关 键 词：合作猪 TRIF基因 生物信息学 表达 

分 类 号：S828.8[农业科学—畜牧学]