节点文献
小麦TaGAPDH5基因的亚细胞定位和表达分析
Subcellular localization and expression analysis of TaGAPDH5 gene in wheat
【摘要】 利用生物信息学方法预测中国春小麦TaGAPDH5的理化性质、亚细胞定位等分子特征,发现该蛋白的相对分子质量为31 541.87,理论等电点(pI)为6.08,为酸性亲水性蛋白,没有信号肽和跨膜结构域.采用基因枪法将TaGAPDH5-GFP融合载体瞬时转化洋葱表皮细胞,发现TaGAPDH5分布于细胞质.通过PLEXdb和Genevestigator数据库下载中国春小麦幼苗、分蘖、拔节等生育期的芯片表达数据,构建TaGAPDH5的芯片表达谱,发现TaGAPDH5在小麦整个生长周期均持续较高水平表达且具有一定组织特异性.采用实时荧光定量PCR(qRT-PCR)技术研究其不同时期的组织表达特性,发现TaGAPDH5从孕穗期到开花期,在同一时期的不同组织中和不同发育时期的同一组织中表达差异较大.通过构建诱导表达谱,发现TaGAPDH5基因能响应多种非生物胁迫.因此推测TaGAPDH5在小麦生长发育过程中发挥重要作用.
【Abstract】 Glyceraldehyde-3-phosphate dehydrogenase(GAPDH) plays pivotal roles in plant metabolism as a ubiquitous protein. However, the precise role of GAPDH in plant is not completely understood. The characteristics of physicochemical properties and subcellular localization of TaGAPDH5 in wheat cv. Chinese spring were analyzed using the bioinformatics software.The acid hydrophilic protein with a molecular weight of 31541.87 and an isoelectric point(pI) of 6.08 have no signal peptide,transmembrane domains. After construction of transient expression vector TaGAPDH5-GFP, particle bombardment with onion epidermal cells indicated that TaGAPDH5 was localized in the cytoplasm. Microarray data of TaGAPDH5 in wheat were downloaded from PLEXdb and Genevestigator with their accession numbers, expression analysis of TaGAPDH5 by microarray revealed that TaGAPDH5 showed remarkable tissue specificity and expressed constantly throughout the plant developmental stages. Quantitative RT-PCR(qRT-PCR) was used to detect organizational expression characteristics of different periods of TaGAPDH5 gene, the result revealed that TaGAPDH5 was differentially expressed between different tissues in the same period,or in the same tissues in different developmental periods. The present study has laid a foundation for subsequently exploring molecular mechanism of regulation of gene expression and signal transmission under drought stress. Relative expression analysis revealed that TaGAPDH5 could be significantly induced by abiotic stresses. Therefore, TaGAPDH5 may play a important role in wheat metabolism.
【Key words】 wheat; TaGAPDH5; subcellular localization; expression analysis;
- 【文献出处】 农业科学研究 ,Journal of Agricultural Sciences , 编辑部邮箱 ,2019年01期
- 【分类号】S512.12
- 【下载频次】482