梅花鹿LHβ基因的克隆及序列分析  

The Cloning and Sequence Analysis of Cervus nippon LHβ Gene

作　　者：竭航[1] 陈彬龙 吴楠[2] 李地艳[2] 雷美艳[1] 张承露[1] 曾德军[1] 

机构地区：[1]重庆市药物种植研究所药用动物研究室,重庆408435 [2]四川农业大学动物遗传育种研究所,四川温江611130

出　　处：《四川农业大学学报》2015年第4期441-446,共6页Journal of Sichuan Agricultural University

基　　金：重庆市基本科研业务费计划项目(2014cstc-jbky-01302;2013cstc-jbky-01325)

摘　　要：【目的】克隆梅花鹿LHβ基因CDS区并进行序列分析。【方法】采用PCR克隆测序的方法获得梅花鹿LHβ基因CDS区全序列,采用ProtParam tool等分子生物学工具对梅花鹿LHβ蛋白的分子量、等电点、二级结构、蛋白功能等进行预测,并采用MEGA 6.0软件构建进化树以确定其系统发育地位。【结果】梅花鹿LHβ基因CDS区全长462bp(GenBank序列号为KT199365),包含了ATG起始密码子和TAA终止密码子,共编码141个氨基酸,蛋白质分子量为15.17kDa,等电点为8.00。梅花鹿LHβ蛋白均位于膜外,无跨膜结构,平均疏水性(GRAVY)为0.391,蛋白功能预测显示,该蛋白最有可能为激素,这与本研究的目的基因相符。进化树分析结果表明,梅花鹿LHβ基因与山羊、绵羊和牛等反刍动物较近,其中与绵羊最近。【结论】梅花鹿LHβ基因CDS区序列与绵羊LHβ基因最近。【Objective】The aim of this study is to clone the LHβgene CDS of Cervus nippon and analyze the structure and features of LHβprotein.【Method】The LHβgene CDS of Cervus nippon was obtained by PCR,cloning and sequencing.Then,the structure and features of LHβprotein,including molecular weight,isoelectric point,secondary structure,function were predicted by online molecular biological tool,such as ProtParam tool.Meanwhile,we constructed a phylogenetic tree by MEGA 6.0software to further explain the genetic basis for this gene.【Results】The full CDS sequence of Cervus nippon LHβ gene was 462 bp（GenBank Accession No.KT199365）,encoding 141 amino acids,which contained ATG start codon,and TGA stop codon,with a calculated molecular weight of 15.17 kDa and an isoelectric point of 8.00.No conserved transmembrane region was detected and all of the amino acids were located outside the membrane.The average of hydrophobicity（GRAVY）was 0.391.Function predicted results showed that the function of this protein most probable act as a hormone,which was in agreement with the target gene.The phylogenetic analysis showed that Cervus nippon had close affiliation with ruminant,including Capra hircus,Ovis aries and Bos taurus,especially Ovis aries.【Conclusion】The LHβgene CDS of Cervus nippon had the closest affiliation with Ovisaries.

关 键 词：梅花鹿 LHΒ基因 克隆 序列分析 

分 类 号：S825[农业科学—畜牧学]