| 邓 利 ，李冬兵，曹玲珑，熊大斌，尹 钧，牛洪斌.大麦衔接蛋白AP 3复合体 HvMu3 基因的克隆及表达分析[J].麦类作物学报,2013,33(3):416 |
| 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 |
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| DOI：10.7606/j.issn.1009-1041.2013.03.02 |
| 中文关键词: 大麦 HvMu3 基因克隆 特征分析 |
| 英文关键词:Barley HvMu3 Gene clone Expression analysis |
| 基金项目: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）。 |
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| 中文摘要: |
| 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4 439 bp，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。 |
| 英文摘要: |
| 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. |
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