遗传 ›› 2011, Vol. 33 ›› Issue (1): 88-94.doi: 10.3724/SP.J.1005.2011.00088

• 研究报告 • 上一篇    

与TaFRA蛋白相互作用候选蛋白的筛选

温小杰, 郝晨阳, 蒲文, 刘旭, 张学勇   

  1. 中国农业科学院作物科学研究所, 国家农作物基因资源与基因改良重大科学工程, 农业部作物种质资源利用重点开放实验室, 北京 100081
  • 收稿日期:2010-08-10 修回日期:2010-09-29 出版日期:2011-01-20 发布日期:2011-01-20
  • 通讯作者: 张学勇 E-mail:xueyongz@ mail.caas.net.cn
  • 基金资助:

    国家863计划项目(编号: 2008AA10Z119)和转基因重大专项(编号: 2009ZX08009-085B)资助。

Screening of the candidate proteins interacting with TaFRA

WEN Xiao-Jie, HAO Chen-Yang, PU Wen, LIU Xu, ZHANG Xue-Yong   

  1. The National Key Facility for Crop Gene Resources and Genetic Improvement, Key Laboratory of Crop Germplasm and Utilization, Min-istry of Agriculture, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2010-08-10 Revised:2010-09-29 Online:2011-01-20 Published:2011-01-20
  • Contact: ZHANG Xue-Yong E-mail:xueyongz@ mail.caas.net.cn

摘要: F-box蛋白是E3泛素连接酶SCF复合体的重要亚基, 通过底物蛋白的特异识别发挥功能。TaFRA是在盐胁迫差异表达片段基础上通过RACE方法获得的一个基因, 编码F-box蛋白。文章利用TaFRA基因构建诱饵表达载体, 直接用cDNA+pGAD+pBD共转化酵母双杂交的方法筛选相互作用的候选蛋白。通过对阳性克隆的鉴定和测序分析, 共获得44个与TaFRA相互作用的候选蛋白, 其中32个为已知蛋白, 包括硫氧还蛋白、金属硫蛋白、ATP合成酶及丝氨酸/苏氨酸蛋白激酶等多种逆境胁迫反应蛋白及转录因子蛋白, 说明TaFRA与胁迫反应相关, 可能通过对上述蛋白编码基因的调控参与了植物的胁迫反应过程, 为进一步阐明TaFRA的功能及作用机制奠定了理论基础。

关键词: 酵母双杂交, 共转化, TaFRA, 相互作用, 候选蛋白

Abstract: F-box protein is an important subunit of SCF complex, an E3 ligase in ubiquitin system, and its function is determined through mediating the specific recognition and combining with substrate protein. TaFRA (F-box protein related to abiotic stress) was identified by RACE based on the fragments diferently expressing in wheat seedling exposed to salt stress and encodes an F-box protein. In this study, pBD-TaFRA bait expression vector was constructed, and cDNA+pGAD+pBD was directly co-transformed into yeast hybrid system to screen condidate proteins interacting with TaFRA. Fourty-four candidate proteins were obtained, in which 32 were known proteins and transcript factors related to stress tolerance such as thioredoxin, metallothinein, ATP synthase, and serine/threonine protein kinase etc. This indicates that TaFRA participates in stress response through regulating above condidate genes, which will provide basis for revealing the mechanism of TaFRA reaction to abiotic stress.

Key words: TaFRA, yeast two hybridization, co-transformation, candidate proteins