藏绵羊生长激素(GH)基因的克隆及序列分析  

Cloning and sequence analysis of Growth Hormone(GH) gene in Tibetan sheep

作　　者：马志杰[1] 魏雅萍[1] 钟金城[2] 马蕊霞[3] 陈雪梅[2] 字向东[2] 陈生梅[1] 

机构地区：[1]青海大学畜牧兽医科学院,青海西宁810016 [2]西南民族大学生命科学与技术学院,四川成都610041 [3]西北农林科技大学动物科技学院,陕西杨陵712100

出　　处：《西北农林科技大学学报（自然科学版）》2008年第7期19-26,共8页Journal of Northwest A&F University(Natural Science Edition)

基　　金：青海省基础研究与软科学计划项目(2006-Z-601)

摘　　要：【目的】对藏绵羊生长激素（GH）基因进行了克隆和序列分析，以期为进一步开展藏绵羊GH基因与其生长发育性状的相关分析以及基因定位、表达调控等研究提供理论基础。[方法】用特定引物对藏绵羊GH基因进行PCR扩增、克降和测序，使用DNAMAN4．0、BioEdit7．0．0、DnaSP4．10．1等生物信息学软件进行序列分析和比较基因组学研究。【结果】藏绵羊GH基因（GenBank登录号EF077162）由5个外显子和4个内含子组成，完整编码序列（Complete coding sequence，CDS）全长为654bp。5个外显子大小分别为13，161，117，162和201bp；4个内含子大小分别为246，227，229和275bp；内含子与外显子的连接区序列遵循基因组成规则。信号肽由26个氨基酸组成，成熟肽由191个氨基酸组成。藏绵羊与已报道的其他绵羊GH基因编码区核苷酸序列及推导的氨基酸序列同源性均为99．0％～100％；而与山羊、普通牛、瘤牛、大额牛、牦牛、水牛各物种在GH基因编码区核苷酸序列上同源性分别为98．4％，98．0％，97．7％，97．5％，97．5％，98．1％；相应的氨基酸序列问同源性分别为100％，99．0％，99．0％，98．1％，98．6％，98．1％。【结论】成功地克隆了藏绵羊GH基因；牛科物种内，藏绵羊与已报道的其他绵羊以及与山羊、普通牛、瘤牛、大额牛、牦牛、水牛等物种在GH基因编码区核苷酸序列和推导的氨基酸序列上均具有较高的保守性。[Objective] The Tibetan sheep growth hormone （GH） gene was cloned and sequenced,comparisons of the sequence and the deduced amino acid sequence with the homologous GH gene and protein sequence of the GHs in other species including other sheep, goats, cattles, zebu, frontalis, yaks and buffalo retrieved from the GenBank were performed in order to provide theoretical foundation for further research of the corelevantion of Tibetan sheep GH gene and growth traits, location and expression i. e. [Method] The Tibetan sheep GH gene was amplified using particular primers,cloned by T-A method and sequenced, based on this the nucleotide and amino acid sequences among different species were compared and analyzed by DNAMAN 4.0,BioEdit 7.0.0,DnaSP 4.10.1 softs. [Result] The numbers of exons and introns of Tibetan sheep GH gene （GenBank Accession No：EF077162） were the same with those in other sheep,goats, cattles,zebu,frontalis,yaks and buffalo, they had 5 exons and 4 introns. The size of CDS sequence of the Tibetan sheep GH gene,encoding 217 amino acids （including signal peptide composed of 26 amino acids and Mature peptide composed of 191 amino acids）, was 654 bp. The size of exon Ⅰ, exon Ⅱ , exonⅢ , exonⅣ and exonVwas 13,161,117,162 and 201 bp respectively;while the size of intron Ⅰ,intronⅡ ,intronⅢ and intron Ⅳ was 246,227,229 and 275 bp respectively. The junction sequence of introns and exons of Tibetan sheep GH gene was in accordance with the law of gene composition. The homology rates of nucleotide sequences of the coding region of GH gene and deduced amino acid sequences of GHs among Tibetan sheep and other sheep were 99%--100%. The homology rates of nucleotide sequences of the coding region of GH gene a- mong Tibetan sheep and goats, cattles, zebu, frontalis, yaks and buffalo were 98. 4%, 98. 0%, 97. 7%, 97.5%,97.5%, 98. 1% respectively,while the homology rates of deduced amino acid sequences of GHs were 100%, 99.0%, 99.0%,98. 1%,98. 6%,98.1% respectively. [Conclusion] The Tibetan sheep G

关 键 词：藏绵羊 生长激素基因 克隆 序列分析 

分 类 号：S826.2[农业科学—畜牧学] Q781[农业科学—畜牧兽医]