| 铁皮石斛两个DEAD-box RNA解旋酶基因克隆及特征分析 |
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| 引用本文：李元敏，李 慧，梁小燕，高 静，沈 霞，颜永刚，李依民，张 岗.铁皮石斛两个DEAD-box RNA解旋酶基因克隆及特征分析[J].西北农业学报,2020,30(7):1059~1067 |
| DOI:10.7606/j.issn.1004-1389.2020.07.011 |
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| 基金项目:陕西中医药大学新进博士科研启动经费（104080001）；陕西中医药大学“秦药”品质评价及资源开发创新团队项目（2019-QN01）；思邈人才专项（141306200105）；陕西省高校首批青年杰出人才支持计划；咸阳市中青年科技领军人才 项目。 |
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| 中文摘要:为获得铁皮石斛DEAD-box RNA解旋酶（RH, RNA Helicase）基因并进行表达及序列分析，利用RACE从其叶片cDNA中分离RH基因，对编码蛋白相关理化特性进行生物信息分析；采用MEGA 6.0构建系统进化树；借助定量PCR技术检测基因表达特性。结果表明：分离到铁皮石斛两个DEAD-box RH基因DoRH1和DoRH2（GenBank注册号KT957555和KT957556），全长为1 648 bp和1 851 bp，各编码1条由428和499个氨基酸组成的肽链，编码蛋白相对分子质量分别为48 206和55 432，等电点5.51和8.65； DoRH1和DoRH2蛋白均包含3个保守的RH结构域和多个基元，无信号肽或跨膜域，预测均定位在细胞核；两个基因与植物DEAD-box RH家族基因相似性高达80%以上；DoRH1、 DoRH2分别聚在拟南芥、水稻、大豆和雷蒙德氏棉DEAD-box RH家族基因进化树的Ⅲ、Ⅳ支；DoRH1和DoRH2表达模式不同，DoRH1在根、茎、叶中表达量无显著差异， DoRH2在根中的相对表达量为叶中的3.26倍，茎与叶中无差异。 |
| 中文关键词:铁皮石斛 RNA解旋酶 基因克隆 表达模式 生物信息学分析 |
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| Cloning and Characterization of Two DEAD-box RNA Helicase Genes in Dendrobium officinale |
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| Abstract:To obtain the DEAD-box RNA Helicase (RH, RNA Helicase) gene from Dendrobium officinale followed by gene expression and sequence analyses, RACE was used to isolate RH genes from the leaf cDNA of D.officinale. Then, the related physicochemical properties of the encoded proteins were analyzed by bioinformatics. MEGA 6.0 was used to build phylogenetic tree for analysing and quantitative PCR technique was emloyed to detect the gene expression profiles. The results showed that the two DEAD-box RH genesDoRH1 andDoRH2 were isolated, 1 648 bp and 1 851 bp in length, respectively (GenBank accessions KT957555 and KT957556).DoRH1 was deduced to a 428 aa (amino acid) protein with a molecular weight of 48 206 and pI of 5.51, whileDoRH2 encoded a 199 aa protein with a molecular weight of 55 432 and pI of 8.65. The two deduced DoRH1 and DoRH2 proteins both had three conserved RH Domains (47-75, 78-251, and 279-424 for the former; 95-123, 126-301, and 329-474 for the latter) and several functional motifs. The two proteins did not have signal peptide or transmembrane domain, and were predicted to locate in the nucleus. The two genes were highly identical to members of DEAD-box RNA helicase family in plant of with similarity of more than 80%. DoRH1 and DoRH2 were respectively grouped into the Ⅲ and Ⅳ branches of the DEAD-box RH family phylogenetic tree from Arabidopsis thaliana, Oryza sativa, Glycine max, and Gossypium raimondii. The expression patterns ofDoRH1 andDoRH2 differed. There was no significant difference for the expression ofDoRH1 in roots, stems, and leaves. The relative expression ofDoRH2 in roots was 3.26 times than that in leaves, but no difference in stems and leaves. |
| keywords:Dendrobium officinale Kimura et Migo RNA helicase Gene cloning Expression pattern Bioinformatics analysis |
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