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FK506结合蛋白(FK506 binding protein,FKBP)具有肽基-脯氨酰顺反异构酶(peptidyl-proly cistrans isomerase,PPIase)活性,可以作为一种分子伴侣在蛋白折叠中发挥作用。本研究采用同源基因克隆获得小麦(Triticum aestivum)高温胁迫应答基因Ta FKBP62c-2B,利用q RT-PCR和绿色荧光蛋白(green fluorescent protein,GFP)示踪技术明确了Ta FKBP62c-2B的表达模式和亚细胞定位信息。序列分析结果表明,小麦Ta FKBP62c-2B基因含有37 bp的5’UTR区、1 926 bp的完整CDS以及176 bp的3’UTR区,共编码642个氨基酸(Gen Bank登录号:KU350629)。结构域分析发现,Ta FKBP62c-2B是由3个FK506结合保守域(FKBP_C)和3个三十四肽重复序列(tetratricopeptide repeat,TPR)构成的多域FKBP。q RT-PCR结果显示,Ta FKBP62c-2B强烈响应高温胁迫;组织表达分析表明,Ta FKBP62c-2B主要在小麦的茎和小穗等植株地上组织中表达,在根部的表达量较低。亚细胞定位表明,Ta FKBP62c-2B位于内质网上。本研究获得了小麦Ta FKBP62c-2B基因全长、明确了其表达规律以及亚细胞定位信息,为进一步了解Ta FKBP62c-2B的生物学功能提供了基础资料。
FK506 binding protein (FKBP) has the activity of peptidyl-proly cistrans isomerase (PPIase), which can act as a chaperone in protein folding. In this study, Ta FKBP62c-2B, a high temperature stress response gene of wheat (Triticum aestivum), was obtained by homologous cloning. The expression pattern of Ta FKBP62c-2B was confirmed by q RT-PCR and green fluorescent protein (GFP) And subcellular localization information. Sequence analysis showed that the Ta FKBP62c-2B gene of wheat contained 37 bp 5 ’UTR region, 1 926 bp complete CDS and 176 bp 3’ UTR region, encoding a total of 642 amino acids (GenBank accession number: KU350629). Domain analysis revealed that Ta FKBP62c-2B is a multi-domain FKBP consisting of three FK506 binding conserved domains (FKBP_C) and three tripeptide peptide repeats (TPRs). q RT-PCR results showed that Ta FKBP62c-2B strongly responded to high temperature stress. Tissue expression analysis showed that Ta FKBP62c-2B was mainly expressed in the shoots and spikelets of wheat plants aboveground tissues, but lower in roots. Subcellular localization showed that Ta FKBP62c-2B is located on the endoplasmic reticulum. The full-length Ta FKBP62c-2B gene of wheat was obtained in this study. The expression pattern and subcellular localization of Ta FKBP62c-2B were confirmed, which provided the basic information for further understanding the biological function of Ta FKBP62c-2B.