羊驼TGF-β1基因的克隆及表达分析  

Cloning and Expression of TGF-β1 Gene from Alpaca(Lama pacos)

作　　者：贾小云[1] 金雷皓 丁娜[1] 罗东萍[2] 范瑞文[3] 董常生[3] 

机构地区：[1]山西农业大学生命科学学院,太谷030801 [2]山西医科大学第一临床医学院风湿免疫科,太原030001 [3]山西农业大学动物科技学院,太谷030801

出　　处：《畜牧兽医学报》2014年第6期885-892,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：教育部高等学校博士学科点专项科研基金(20111403120006);中国博士后科学基金普通资助和特别资助(20100481306;2012T50246);博士后经费(83557);山西省留学基金(2011051);山西人才引进与开发专项(614191);山西省高等学校131人才;山西农业大学中青年学术骨干

摘　　要：旨在克隆羊驼TGF-β1基因,了解TGF-β1序列特征及其在不同组织中的表达特征,寻找TGF-β1基因的mRNA表达量与毛色之间的相关性,为研究TGF-β1基因的生物学功能提供基础。以羊驼为材料,利用3′和5′RACE技术克隆羊驼TGF-β1基因,利用DNAman、PROSITE和megAlign软件分别分析序列特征和相似性,应用荧光定量PCR(qRT-PCR)分析TGF-β1基因在不同被毛颜色羊驼皮肤组织中的表达及TGF-β1在山羊不同组织中的表达特异性。结果表明,羊驼TGF-β1cDNA克隆全长为1 364bp(GenBank登录号:KF887499),ORF长1 173bp,编码390个氨基酸。氨基酸序列分析显示,羊驼TGF-β1具有其家族的典型结构特征,与其他哺乳动物相似性很高,与野猪的亲缘关系最近。羊驼TGF-β1基因在棕色羊驼皮肤组织中的表达量是白色羊驼皮肤组织的3.5倍。组织特异性分析表明,TGF-β1在山羊不同组织中均有表达,且在淋巴组织中表达量相对最高。羊驼TGF-β1全长cDNA的克隆和不同组织器官TGF-β1基因表达分析显示,TGF-β1与羊驼毛色形成密切相关。本研究为深入探讨TGF-β1参与动物毛色形成及调控的分子机制奠定了基础。In order to explore the biological functions of TGF-β1 in coat color formation in alpaca,the experiment was conducted to clone alpaca TGF-β1 gene,investigate the expression of TGF-β1 in different tissues/organs,and examine the relationship between the TGF-β1 expression and alpaca coat color.The TGF-β1cDNA was cloned from alpaca by 3′ and 5′RACE.The sequence analysis was performed by DNAman,PROSITE and megAlign.The expression levels of TGF-β1 gene in different organs/tissues were analyzed by qRT-PCR.The full length of TGF-β1cDNA in alpaca was 1 364bp(GenBank Accession No.KF887499)with 1 173bp ORF coding 390 amino acids.Sequence analysis showed alpaca TGF-β1 has typical structures of TGF-β1 family,showing high similarity with responsed proteins from other mammals.Phylogenetic analysis showed that alpaca TGF-β1 gene is much closed to wild boar(Sus scrofa).qRT-PCR showed that the relativegene expression level of TGF-β1 in brown alpaca was 3.5 times than that in white alpaca.TGF-β1 gene was expressed widely in tested tissues in goat while the expression levels were different in those tissues and highest in lymphoid tissue.Cloning and sequence characterization of alpaca TGF-β1 gene plus the expression analysis in different tisssues/organs indicate that TGF-β1 may play important role in alpaca coat color formation,provide the related DNA elements and experimental basic for further investigating on the molecular mechanism of TGF-β1 functions in animal coat color formation and regulation.

关 键 词：羊驼 TGF-Β1基因 CDNA末端快速扩增 QRT-PCR 表达分析 

分 类 号：S829.9[农业科学—畜牧学]