胡桃楸MYB17基因克隆及功能分析  

Cloning and Functional Analysis of MYB17 Gene in Juglans mandshurica

作　　者：杨琦 高瑞馨[1] Yang Qi;Gao Ruixin(College of Life Science,Northeast Forestry University,Harbin,150040)

机构地区：[1]东北林业大学生命科学学院,哈尔滨150040

出　　处：《分子植物育种》2022年第18期5995-6000,共6页Molecular Plant Breeding

基　　金：国家科技基础资源调查专项(No.2019FY100500)资助。

摘　　要：本研究旨在探究一个受干旱诱导的胡桃楸MYB17基因的序列特征。以两年生胡桃楸叶片为材料,提取RNA,反转录为cDNA,通过胡桃楸转录组相关信息,用RT-PCR技术克隆得到胡桃楸MYB17基因。经生物信息学分析,胡桃楸MYB17基因全长951 bp,共编码316个氨基酸;胡桃楸MYB17基因等电点为5.61,相对分子质量为35.10 kD。进化关系分析表明,胡桃楸MYB17与胡桃MYB17基因有密切的遗传关系。分析了胡桃楸MYB17蛋白的胞内定位、二级结构、三级结构和结构域。将胡桃楸MYB17基因构建到酿酒酵母表达载体上进行抗性分析,发现对比阴性对照,转MYB17基因的酵母在干旱胁迫、盐胁迫和碱胁迫中的抗性都有不同程度的提高。本研究发现的胡桃楸MYB17蛋白的理化性质及其功能分析为进一步研究胡桃楸的生物学功能提供了理论依据。The pur pose of this study was to investigate the sequence characteristics of MYB17 gene in a drought-induced Juglans mandshurica.RNA was extracted from the leaves of the two-year Juglans sativus,which was reverse-transcriptional to cDNA.Through the transcriptome information of Juglans mandshurica,MYB17 gene was cloned by RT-PCR and named MYB17 gene.Bioinformatics analysis showed that the entire population of MYB17gene was 951 bp,encoding a total of 316 amino acids.The isoelectric point of MYB17 gene was 5.61 and its relative molecular weight was 35.10 kD.Analysis of evolutionary relationship showed that there was a close genetic relationship between Juglans mandshurica MYB17 and Walnut MYB17 gene.The intracellular location,secondary structure,tertiary structure and domain of MYB17 were analyzed.The MYB17 gene was constructed into Saccharomyces cerevisiae expression vector for resistance analysis,and it was found that compared with the negative control,the resistance of MYB17 transgenic yeast in drought stress,salt stress and alkali stress were improved to varying degrees.The physicochemical properties and functional analysis of the MYB17 protein of Juglans mandshurica in this study provide a theoretical basis for further research on the biological functions of Juglans mandshurica.

关 键 词：胡桃楸 MYB17蛋白 基因克隆 生物信息学分析 胁迫 

分 类 号：S792.132[农业科学—林木遗传育种]