甘蔗ATP结合蛋白基因克隆及其干旱胁迫下的表达特性分析

发布时间：2018-01-23 04:13

本文关键词： 甘蔗 ATP结合蛋白基因克隆生物信息学干旱胁迫表达特性　出处：《南方农业学报》2017年09期 　论文类型：期刊论文

【摘要】：【目的】克隆甘蔗ATP结合蛋白基因,分析其基本生物学信息及在干旱胁迫下的表达特性,为ATP结合蛋白的功能研究提供理论依据。【方法】从乙烯诱导甘蔗差异表达转录组和水分胁迫下甘蔗cDNA文库中获取ATP结合蛋白基因序列,设计其引物进行RT-PCR扩增,利用生物信息学软件进行序列分析及其蛋白结构和功能预测,并采用实时荧光定量PCR(qPCR)检测干旱胁迫处理下的甘蔗ATP结合蛋白基因的表达情况。【结果】克隆获得的甘蔗ATP结合蛋白基因全长2145 bp,编码714个氨基酸,其蛋白分子质量为79.430 kD,理论等电点(pI)为9.33,不稳定系数为44.38;甘蔗ATP结合蛋白与玉米ATP结合蛋白的核苷酸序列及其推导氨基酸系列同源性最高,均达93%。甘蔗ATP结合蛋白主要由无规则卷曲(42.02%)、α螺旋(25.63%)和延伸链(22.27%)结构组成,具有蛋白激酶催化结构域、ATP结合位点、核苷酸结合位点(NBS)、糖基化位点、激酶磷酸化结合位点及核定位信号等;甘蔗ATP结合蛋白可能主要定位在细胞核、叶绿体、线粒体和过氧化物酶体中,起中间代谢调控作用,或参与转录调控、免疫应答、胁迫应答及信号转导等生物反应。qPCR检测结果显示,在干旱胁迫7 d后,甘蔗ATP结合蛋白基因的表达量明显下调,约为对照的30%,恢复正常供水(复水)后,表达量回升。【结论】甘蔗ATP结合蛋白基因表达受干旱胁迫诱导,可能参与甘蔗对干旱逆境胁迫响应的代谢调控。
[Abstract]:[objective] to clone sugarcane ATP binding protein gene and analyze its basic biological information and expression characteristics under drought stress. [methods] the sequence of ATP binding protein gene was obtained from the differential expression transcriptome of sugarcane induced by ethylene and the cDNA library of sugarcane under water stress. The primers were designed for RT-PCR amplification, sequence analysis and protein structure and function prediction were carried out by bioinformatics software. The real-time fluorescence quantitative PCR- qPCR was used. The expression of ATP binding protein gene in sugarcane under drought stress was detected. [results] the ATP binding protein gene of sugarcane was 2145bp in length. It encodes 714 amino acids with molecular weight of 79.430 KD, theoretical isoelectric point (Pi) of 9.33 and instability coefficient of 44.38; The nucleotide sequence and deduced amino acid sequence of sugarcane ATP binding protein and maize ATP binding protein have the highest homology. The ATP binding protein of sugarcane was mainly composed of 42.022KT, a helix 25.63 and 22.27m). There are ATP binding sites, nucleotide binding sites, glycosylation sites, kinase phosphorylation binding sites and nuclear localization signals. The ATP binding proteins in sugarcane may be located in the nucleus chloroplast mitochondria and peroxisome which play an intermediate role in metabolic regulation or participate in transcriptional regulation and immune response. The results of stress response and signal transduction. QPCR showed that after 7 days of drought stress, the expression of ATP binding protein gene in sugarcane was significantly down-regulated, about 30% of the control. [conclusion] the expression of ATP binding protein gene in sugarcane was induced by drought stress, which may be involved in the metabolic regulation of sugarcane response to drought stress.
【作者单位】：广西作物遗传改良生物技术重点开放实验室;广西农业科学院甘蔗研究所/农业部广西甘蔗生物技术与遗传改良重点实验室/广西甘蔗遗传改良重点实验室;
【基金】：广西自然科学基金项目(2013GXNSFAA019077,2013GXNSFAA019088,2014GXNSFAA118081) 广西八桂学者专项项目([2013]3号)
【分类号】：S566.1
【正文快照】： (1Guangxi Crop Genetic Improvement and Biotechnology Laboratory,Nanning 530007,China;2Sugarcane ResearchInstitute,Guangxi Academy of Agricultural Sciences/Key Laboratory of Sugarcane Biotechnology and GeneticsImprovement(Guangxi),Ministry of Agriculture/

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