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期 刊 ：植物遗传资源学报 2014年 15卷 1期

关键词：小麦；UCE2；表达模式；酵母双杂交；蛋白纯化；

Keywords:wheat, UCE2, expression pattern, yeast-two-hybrid system, protein purification,

摘 要 ：通过酵母双杂交系统筛选小麦抗逆相关蛋白TaMAPK2的互作蛋白，获得候选蛋白TaE2。生物信息学分析表明，TaE2基因属于泛素结合酶UCE2基因家族成员。半定量RT-PCR结果表明TaE2基因在小麦根、茎、叶和种子等器官中均有表达，荧光定量PCR显示TaE2基因在干旱、高盐和ABA胁迫下均上调表达。利用原核表达获得GST-E2融合蛋白并使用蛋白标记亲和层析柱纯化。本文报道的小麦TaE2基因，为进一步分析泛素蛋白酶体途径在小麦逆境响应中的功能和机制打下基础。

Abstract:The ubiquitin-conjugating enzyme, named TaE2, was obtained as the candidate interaction protein of TaMAPK2 by yeast-two-hybrid system. Bioinformatic analysis showed that TaE2 was a member of ubiquitin-conjugating enzyme E2 family. Semi-quantitative RT-PCR showed that TaE2 was expressed in root, stem, leaves and seeds. TaE2 was up-regulated under drought, high-salt and ABA treatments. The fusion protein GST-TaE2 was expressed in Escherichia.coli and purified by GST-Trap HP. TaE2 gene reported in this study lay a foundation for further studying the function and mechanisms of the ubiquitin proteasome pathway response to stresses.

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