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2025年4月3日 星期四
  2016, Vol. 24 Issue (7): 997-1007    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
小麦TaFKBP62c-2B基因克隆、组织表达及亚细胞定位
吴迪1,陈林2,张向展2,李嵩2,李敏2,刘子辉2,孙希媛2,郑炜君1,柴守诚1
1. 西北农林科技大学
2. 西北农林科技大学 农学院
Cloning, Expression and Subcellular Localization of TaFKBP62c-2B Gene in Wheat (Triticum aestivum)
1, 1, 1, 1, 1, 1, 1, 1
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摘要 FK506结合蛋白(FK506 binding protein, FKBP)具有肽基-脯氨酰顺反异构酶 (peptidyl-proly cis-trans 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TaFKBP62c-2B,利用qRT-PCR和绿色荧光蛋白(green fluorescent protein, GFP)示踪技术明确了TaFKBP62c-2B的表达模式和亚细胞定位信息。序列分析结果表明,小麦TaFKBP62c-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(GenBank登录号: KU350629)。结构域分析发现,TaFKBP62c-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。qRT-PCR结果显示,TaFKBP62c-2B强烈响应高温胁迫;组织表达分析表明,TaFKBP62c-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,TaFKBP62c-2B位于内质网上。本研究获得了小麦TaFKBP62c-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TaFKBP62c-2B的生物学功能提供了基础资料。
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作者相关文章
吴迪
陈林
张向展
李嵩
李敏
刘子辉
孙希媛
郑炜君
柴守诚
关键词 小麦FK506结合蛋白(FKBP)高温胁迫表达分析亚细胞定位    
Abstract:FK506-binding proteins (FKBPs) are known as the receptor for the immunosuppressant drug FK506. FKBPs are ubiquitously distributed in all living organisms and highly conserved during evolution. FKBPs have peptidyl-proly cis-trans isomerase (PPIase) activity and can be used as molecular chaperone to have a function in protein folding. Plant FKBPs take part in diversity of cellular processes including plant growth and abiotic stress responses. In this study, TaFKBP62c-2B was isolated from wheat (Triticum aestivum) by RT-PCR and the expression pattern of this gene was analyzed through qRT-PCR. The subcellular localization of TaFKBP62c-2B was also investigated by using the green fluorescent protein (GFP) method. The sequencing analysis showed that TaFKBP62c-2B gene consisted of 1 926 bp CDS, 37 bp 5' UTR and 176 bp 3' UTR, and encoded 642 amino acids (GenBank accession No: KU350629). The amino acid sequence analysis demonstrated that TaFKBP62c-2B had 3 conserved FK506 binding domains (FKBd), which were respectively named 62c-2BⅠ, 62c-2BⅡand 62c-2Ⅲ, and 3 tetratricopeptide repeats (TPR), so TaFKBP62c-2B belonged to multiple-domain FKBPs. The amino acid alignment analysis showed that 62c-2BⅠ was the most similar conserved domain with HsFKBP12a and should be the key domain for the PPIase function of TaFKBP62c. The 9 homologus genes from 9 different species of Aegilops tauschii, Triticum urartu, millet (Setaria italic), barley (Hordeum vulgare), Brachypodium distachyon, sorghum (Sorghum bicolor), maize (Zea mays), soybean (Glycine max), rice (Oryza sativa indica Group) of TaFKBP62c-2B were searched and found in NCBI database and the identity of amino acid between 9 homologous genes and TaFKBP62c-2B ranged of 86%~96%. The highest identity of amino acid was between EMS49100.1(T. urartu) and TaFKBP62c-2B, while the lowest was between soybean (XP_014634970.1) and TaFKBP62c-2B. Then phylogenetic tree of FKBP62c was constructed using these 9 genes and TaFKBP62c-2B. The expression pattern disclosed the expression level of TaFKBP62c-2B was increased immediately under heat stress, it came to the peaks at 2 h and droped gradully after that. TaFKBP62c-2B was responded to drought stress, the expression came to the peaks at 1 h and droped, it came to another peak at 8 h. Tissue expression pattern analysis showed that TaFKBP62c-2B expressed in multiple tissues. Stem accumulated higher TaFKBP62c-2B transcription compared with the spike, spikelet, lemma and palea, while root and leaf had the least TaFKBP62c-2B transcription among all the tissues. According to the prediction database, TaFKBP62c-2B was localized to endoplasmic reticulum (ER). The subcellular location were validated by means of fusing TaFKBP62c-2B with N-terminal GFP and then expressed in wheat protoplats using PEG transfection method. The green fluorescence were mainly observed in cytoplasm, and were not seen in chloroplasts. TaFKBP62c-2B should be localized to ER. In this study, the TaFKBP62c-2B gene was isolated, which the expression pattern and subcellular localization were detected. All above results pave the way to further function analysis of TaFKBP62c-2B.
Key wordsWheat    FK506 binding protein (FKBP)    Heat stress    Expression analysis    Subcellular localization
收稿日期: 2016-01-13      出版日期: 2016-05-20
ZTFLH:  S332.5  
基金资助:国家科技支撑计划项目;陕西省科技统筹创新项目;西北农林科技大学博士启动基金
通讯作者: 柴守诚     E-mail: chaishoucheng@126.com
引用本文:   
吴迪 陈林 张向展 李嵩 李敏 刘子辉 孙希媛 郑炜君 柴守诚. 小麦TaFKBP62c-2B基因克隆、组织表达及亚细胞定位[J]. , 2016, 24(7): 997-1007.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2016/V24/I7/997
 
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