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Molecular cloning and characterization of a gene encoding RING zinc finger ankyrin protein from drought-tolerant Artemisia desertorum

机译:耐旱蒿蒿环RING锌指锚蛋白蛋白编码基因的分子克隆与鉴定

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摘要

A RING zinc finger ankyrin protein gene, designated AdZFPl, was isolated from drought-tolerant Artemisia desertorum Spreng by mRNA differential display and RACE. Its cDNA was 1723 bp and encoded a putative protein of 445 amino acids with a predicted molecular mass of 47.9 kDa and an isoelectric point (pI) of 7.49. Atypical C3HC4-type RING finger domain was found at the C-terminal region of the AdZFP1 protein, and several groups of ankyrin repeats were found at the N-terminal region. Alignments of amino acid sequence showed that AdZFP1 was 66% identical to the Arabidopsis thaliana putative RING zinc finger ankyrin protein AAN31869. Transcriptional analysis showed that AdZFP1 was inducible under drought stress in root, stem and leaf of the plant. Semi-quantitative reverse-transcriptase-polymerase chain reaction (RT-PCR) analysis showed that the transcript of AdZFP1 was strongly induced by exogenous abscisic acid (ABA) and also by salinity, cold and heat to some extent. Overexpression of the AdZFP1 gene in transgenic tobacco enhanced their tolerance to drought stress.
机译:通过mRNA差异显示和RACE从耐旱的沙蒿蒿中分离出命名为AdZFP1的RING锌指锚蛋白蛋白基因。它的cDNA为1723 bp,编码一个445个氨基酸的推定蛋白质,预测分子量为47.9 kDa,等电点(pI)为7.49。在AdZFP1蛋白的C末端区域发现了非典型的C3HC4型RING指结构域,在N末端区域发现了几组锚蛋白重复序列​​。氨基酸序列的比对表明,AdZFP1与拟南芥推定的RING锌指锚蛋白锚蛋白AAN31869具有66%的同一性。转录分析表明,AdZFP1在干旱胁迫下可在植物的根,茎和叶中诱导。半定量逆转录聚合酶链反应(RT-PCR)分析表明,AdZFP1的转录被外源脱落酸(ABA)强烈地诱导,并且在一定程度上受盐度,冷热影响。转基因烟草中AdZFP1基因的过表达增强了其对干旱胁迫的耐受性。

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