合作猪IFN-ε基因克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of IFN-ε Gene in Hezuo Pigs

作　　者：杨姣姣[1] 谢开会 王伟 闫尊强 杨巧丽 马艳萍[2] 滚双宝[1,3] YANG Jiaojiao;XIE Kaihui;WANG Wei;YAN Zunqiang;YANG Qiaoli;MA Yanping;GUN Shuangbao(College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;Library of 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]甘肃农业大学图书馆,兰州730070 [3]甘肃省现代养猪工程技术研究中心,兰州730070

出　　处：《西北农业学报》2020年第1期11-20,共10页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金(31660646);甘肃农业大学人才专项经费(2017RCZX-15)~~

摘　　要：旨在克隆合作猪干扰素ε(Interferon epsilon,IFN-ε)基因,并对其结构和编码蛋白进行生物信息学分析,为进一步研究合作猪IFN-ε生物学功能提供参考。根据GenBank上公布的猪IFN-ε基因序列(登录号:NM_001105310.1)设计引物,PCR扩增及测序获得合作猪IFN-ε基因完整编码区序列(Coding sequence,CDS),并对其进行生物信息学分析。结果显示,合作猪IFN-ε基因编码区长582 bp,编码193个氨基酸。该基因编码区序列与杜洛克猪、人、小鼠、黑猩猩、狗、瘤牛、蒙古野马、山羊和绵羊的同源性分别为98.4%、77.7%、56.8%、78.2%、76.5%、87.0%、85.5%、85.0%和86.0%;系统进化树结果表明,合作猪与瘤牛、蒙古野马、绵羊、山羊亲缘关系较近。此外,在合作猪IFN-ε基因CDS上共发现3个碱基突变,均为错义突变。氨基酸序列分析表明,合作猪IFN-ε蛋白分子式为C 1027 H 1630 N 278 O 290 S 10,理论分子质量为22.83 ku,等电点(pI)为8.43,属于碱性蛋白;平均亲水系数为-0.240,属于亲水蛋白质;IFN-ε蛋白无跨膜结构,存在信号肽序列,属于分泌型蛋白;IFN-ε蛋白只包含1个超家族保守结构域,即IFab结构域。二级结构分析表明,该蛋白中α-螺旋、无规则卷曲、β-转角和延伸链所占比例分别为61.66%、30.05%、3.11%和5.18%。This study aimes at the structure and encoded protein of the interferon epsilon(IFN-ε)gene in Hezuo pig by clone and bioinformatic methods,which will provide a reference for further studyon the biological function of IFN-εin Hezuo pig.The primers were designed based on the porcine IFN-εgene reference sequence in GenBank(accession number:NM_001105310.1).Additionally,the complete coding sequence(CDS)of the porcine IFN-εgene was obtained by PCR amplification and sequencing,and then was analyzed byusing bioinformatics.The results showed that the coding region of IFN-εgene in Hezuo pig was 582 bp in length and encoded 193 amino acids.The homology analysis found that coding region sequence of IFN-εin Hezuo pighad similarity of 98.4%,77.7%,56.8%,78.2%,76.5%,87.0%,85.5%,85.0%and 86.0%compared with Duroc,human,mouse,chimpanzee,dog,zebu,Equus przewalskii,goats and sheep,respectively;the result of phylogenetic tree showed that IFN-εgene in Hezuo pigs was closely related to zebu,Equus przewalskii,sheep and goats.In addition,a total ofthree bases of IFN-εgene CDS shappened missense mutations.Amino acid sequence analysis showed that the molecular formula,theoretical molecular mass,and isoelectric point of IFN-εprotein were C 1027 H 1630 N 278 O 290 S 10,22.83 ku and 8.43,respectively.The IFN-εprotein was the hydrophilic and basic protein with total average hydrophilic index-0.240;IFN-εprotein was a secretory protein and had a signal peptide sequence without membrane structure;IFN-εprotein contained one superfamily conserved domain,the IFab domain.The secondary structure analysis of IFN-εprotein showed that the percent of amino acids ofα-helix,random coil,β-turn and extended chain were 61.66%,30.05%,3.11%and 5.18%,respectively.This study will provide the basis for further research on the function of IFN-εin Hezuo pigs.

关 键 词：合作猪 IFN-ε基因 CDS 克隆 生物信息学分析 

分 类 号：S828[农业科学—畜牧学]