小麦泛素结合酶TaE2的表达分析及蛋白互作

Expression Pattern and Protein Interaction Analysis of a Wheat Ubiquitin-conjugating Enzyme TaE2

以小麦抗逆相关蛋白TaMAPK2作为诱饵,利用酵母双杂交系统筛选小麦cDNA融合表达文库,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。

The ubiquitin-conjugating enzyme, named TaE2, was obtained as the interaction protein of wheat stress-related TaMAPK2,which was used as bait protein to screen the wheat cDNA library 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.