大麦衔接蛋白AP-3复合体HvMu3基因的克隆及表达分析  

Cloning and Expression Analysis of the Subunit HvMu3 of Adaptor Complexes AP-3 from Barley

作　　者：邓利[1] 李冬兵[1] 曹玲珑[1] 熊大斌[1] 尹钧[1] 牛洪斌[1] 

机构地区：[1]河南农业大学国家小麦工程技术研究中心,河南郑州450002

出　　处：《麦类作物学报》2013年第3期416-422,共7页Journal of Triticeae Crops

基　　金：农业部转基因生物新品种培育重大专项(2011ZX08002-003);科技部"863"计划分子育种项目(2012AA101105)

摘　　要：为研究大麦衔接蛋白AP-3复合体的功能,根据Mu3亚基的保守氨基酸序列,借助生物信息学搜索大麦EST序列,拼接了一个大麦(Hordeum vulgare)衔接蛋白AP-3复合体Mu3亚基基因(HvMu3),以大麦叶片来源的DNA做为模板,采用RT-PCR扩增HvMu3基因。采用半定量RT-PCR及荧光定量PCR(qRT-PCR)对该基因的组织表达特性以及ABA、NaCl和Cr6+胁迫条件下的表达模式进行分析。结果表明,HvMu3完整阅读框全长为4 439bp,包含8外显子和7内含子,编码417个氨基酸残基,其中含有1个MHD结构域,属于衔接蛋白AP-3复合体Mu3亚基的典型特征,是首次克隆获得大麦衔接蛋白AP-3复合体Mu3亚基基因HvMu3;HvMu3在供试样品的根、茎、叶、胚和胚乳中均能表达,以胚中表达量最高,根、茎和叶次之,胚乳中表达量最低,表明该基因在大麦的旺盛生长组织中表达强烈;HvMu3在大麦幼苗叶片中的表达受ABA、NaCl和Cr6+胁迫诱导,推测其在大麦的生长发育和抵抗逆境胁迫过程中发挥重要作用。In this study the μ3 subunit gene HvMu3,subunit μ3 of complexes AP3 from barley（Hordeum vulgare） was isolated with RT-PCR using a pair of primers designed based on simulation of gene splicing and the conserved amino acid sequence of the Mu3 subunits.Semi-quantitative RT-PCR and q-PCR assay were performed to examine tissue expression of HvMu3 during NaCl,ABA and Cr6＋ stress treatments.The results showed that：（1） HvMu3 contained a 4 439 bp long complete open reading frame including 8 exons and 7 introns,and encoded 417 amino acid residues containing a MHD domain,the typical characteristics of Mu3 3 subunits.（2） As displayed by semi-quantitative RT-PCR and q-PCR assay,HvMu3 was expressed in all tissues,and its expression level was high in the embryo and root,suggesting a close relationship between the expression of this gene and the development of barley.（3） The expression profiling also showed that HvMu3 was up-regulated by ABA,NaCl and Cr6＋ stresses in the seedlings.

关 键 词：大麦 HvMu3 基因克隆 特征分析 

分 类 号：S512.3[农业科学—作物学] S330