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刚毛柽柳ThGAI基因的克隆和序列分析

发表时间:2017-08-15  浏览量:45  下载量:8
全部作者: 张悦,张玉,张春蕊,王超
作者单位: 东北林业大学林木遗传育种国家重点实验室
摘 要: 通过对刚毛柽柳转录组的分析,克隆获得了一条柽柳DELLA家族蛋白基因,命名为ThGAI. 序列分析结果表明,ThGAI基因全长cDNA为2 901 bp,包括1 980 bp的开放阅读框(open reading frame,ORF),推测ThGAI基因编码659个氨基酸,编码蛋白相对分子质量为71.99 ku,等电点为5.58. 通过生物信息学分析发现,ThGAI基因编码蛋白具有DELLA蛋白特有的结构域;氨基酸多序列比对和系统进化树分析发现,ThGAI基因编码氨基酸序列与其他已知植物DELLA蛋白有较高的同源性,属于DELLA亚家族成员,并具有TVHYNP结构域、POLY S/T/V结构域、核定位NLS、VHVID结构域、亮氨酸结构域及GRAS结构域,且与玉米ZmGAI亲缘性最近。实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)分析表明,ThGAI表达受NaCl和聚乙二醇(PEG)诱导,暗示赤霉素(gibberellins,GA)可能参与了刚毛柽柳的非生物胁迫响应过程。
关 键 词: 分子生物学;刚毛柽柳;ThGAI基因;DELLA蛋白;表达分析
Title: Gene cloning and expression analysis of ThGAI from Tamarix hispida
Author: ZHANG Yue, ZHANG Yu, ZHANG Chunrui, WANG Chao
Organization: State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University
Abstract: A protein gene of DELLA family named ThGAI gene was successfully cloned from the transcrip cDNA library of Tamarix hispida. Sequence analysis indicated that ThGAI gene is 2 901 bp in length, containing an open reading frame (ORF) of 1 980 bp and encoding 659 amino acids. The estimated molecular weight and isoelectric point of the protein encoded were 71.99 ku and 5.58, respectively. Bioinformatics analysis indicated that the typical domains of DELLA protein were found in the protein encoded by ThGAI gene. Amino acid sequence alignment with other species and phylogenetic tree analysis indicated that the deduced amino acid sequence of ThGAI gene was highly homologous with those of DELLA protein from other plants and was belonged to the DELLA family. They had the DELLA protein typical domains such as TVHYNP, POLY S/T/V, NLS, VHIID, leucine and the conserved GRAS. They had the highest homology with ZmGAI from Zea mays Linn. Quantitative real-time PCR (qRT-PCR) assay revealed that the mRNA levels of ThGAI was significantly up-regulated under NaCl and PEG treatments, suggesting that gibberellins (GA) may be involved in abiotic stress response of T. hispida.
Key words: molecular biology; Tamarix hispida; ThGAI gene; DELLA protein; expression analysis
发表期数: 2017年8月第15期
引用格式: 张悦,张玉,张春蕊,等. 刚毛柽柳ThGAI基因的克隆和序列分析[J]. 中国科技论文在线精品论文,2017,10(15):1679-1687.
 
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