木薯MeAHL17基因的克隆及表达分析OA北大核心CSTPCD
Cloning and Expression Analysis of MeAHL17 Gene in Cassava
AT-hook核定位蛋白(AT-hook nuclear localized proteins,AHLs)是一种小的DNA结合蛋白基序,在植物的生长发育、器官构建、逆境胁迫与激素信号应答等方面发挥重要作用.本研究从华南8号木薯(SC8)中克隆获得MeAHL17基因,利用生物信息学方法对MeAHL1 7基因进行启动子分析、蛋白理化性质分析、保守功能域的预测和序列比对.结果表明:MeAHL17基因的开放阅读框长度为939 bp,编码一个具有312个氨基酸的蛋白,其理论等电点为6.89,分子式为C1420H2215N415O451S11,分子量为32.669 38 kDa,带正电荷氨基酸残基总数(Lys+Arg)为23,带负电荷氨基酸残基总数(Asp+Glu)为24,脂肪系数为57.47,总平均亲水性系数(GRAVY)为-0.491,不稳定系数为58.06;该蛋白不含有信号肽,位于细胞膜内,无跨膜结构,含有5个糖基化位点和45个磷酸化位点,是一个不稳定的亲水性酸性蛋白;MeAHL17蛋白包含AT-hook保守核心序列和PPC结构域的保守核心序列,符合AT-hook家族的典型结构特征.通过亚细胞定位和转录活性分析实验证明MeAHL17是一个定位于细胞核且具有转录活性的转录因子.组织表达模式分析表明,MeAHL1 7基因主要在体胚、须根和愈伤组织中表达.不同激素处理表明,MeAHL1 7基因受到乙烯(ACC)、茉莉酸甲酯(JA)和生长素(IAA)等激素的诱导表达,推测其可能参与了乙烯、茉莉酸甲酯和生长素信号途径.非生物胁迫处理表明,MeAHL17基因响应干旱和盐胁迫.本研究初步确定了木薯MeAHL17基因在生长发育、激素信号和逆境胁迫等方面具有重要的作用,为进一步研究其功能提供理论依据和参考.
AT-hook nuclear localized proteins(AHLs)are small DNA-binding protein motifs that play an important role in plant growth and development,organ construction,stress and hormone signaling response.In this study,MeAHL17 was cloned from cassava cultivar SC8.Bioinformatics methods were used to analyze MeAHL17 promoter,protein phys-icochemical properties,prediction of conserved functional domain and sequence comparison.The results showed that the open reading frame length of MeAHL17 gene was 939 bp,encoding a protein with 312 amino acids,its theoretical isoelectric point was 6.89,its molecular formula was C1420H2215N415O451S11,and its molecular weight was 32.669 38 kDa.The total number of positively charged amino acid residues(Lys+Arg)was 23,the total number of negatively charged amino acid residues(Asp+Glu)was 24,the fat coefficient was 57.47,the total average hydrophilic coefficient(GRAVY)was-0.491 and the instability coefficient was 58.06.The protein contains no signal peptide,is located in the cell mem-brane,has no transmembrane structure,contains 5 glycosylation sites and 45 phosphorylation sites,and is an unstable hydrophilic acidic protein MeAHL17 contained AT-hook conserved core sequences and PPC domain conserved core sequences,which was consistent with the typical structural characteristics of AT-hook family.Through subcellular lo-calization and transcriptional activity analysis experiments,MeAHL17 was proved to be a nucleus localized transcrip-tion factor with transcriptional activity.Tissue expression pattern analysis showed that MeAHL17 was mainly expressed in somatic embryo,fibrous root and callus.Different hormone treatments showed that MeAHL17 was induced by ethyl-ene(ACC),methyl jasmonate(JA)and growth hormone(IAA),which suggested that MeAHL17 might be involved in ethylene,methyl jasmonate and growth hormone signaling pathway.Abiotic stress treatment showed that MeAHL17 was responsive to drought and salt stress.This study preliminarily identified the important role of MeAHL17 in growth and development,hormone signaling and stress,and provided theoretical basis and reference for further study of its function.
张亚文;王晓彤;张兴龙;唐湘宁;刘姣;郭建春
海南大学生命健康学院,海南海口 570228||中国热带农业科学院热带生物技术研究所/农业农村部热带作物生物学与遗传资源利用重点实验室/海南热带农业资源研究院/海南省热带农业生物资源保护与利用重点实验室,海南海口 571101||中国热带农业科学院三亚研究院,海南三亚 572000中国热带农业科学院热带生物技术研究所/农业农村部热带作物生物学与遗传资源利用重点实验室/海南热带农业资源研究院/海南省热带农业生物资源保护与利用重点实验室,海南海口 571101||中国热带农业科学院三亚研究院,海南三亚 572000
农业科学
木薯MeAHL17基因克隆亚细胞定位转录活性分析表达模式分析
cassavaMeAHL17gene cloningsubcellular localizationtranscriptional activity analysisexpression pat-tern analysis
《热带作物学报》 2024 (007)
1303-1313 / 11
国家自然科学基金项目(No.32001602);海南省自然科学基金项目(No.323MS087);现代农业产业技术体系建设专项(No.CARS-11-HNGJC).
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