拟南芥 AtTR1 在盐胁迫应答中的功能初探

本研究主要探索油菜中E3泛素连接酶BnTR1在拟南芥中的同源基因AtTR1(At3g47550)的功能.通过体外泛素化实验证明AtTR1具有E3连接酶活性.基因表达分析显示该基因受200mmol/L NaCl显著诱导,说明该基因可能在响应盐胁迫中发挥一定的功能.为了更深入的探究该基因在植物耐盐中的作用,构建了植物表达载体pZH01-AtTR1转化突变体.在含有潮霉素的培养基上筛选阳性苗,并利用荧光定量PCR检测表明AtTR1基因已经成功转入突变体中.

First exploration on Protein Function of Arabidopsis AtTR1 in response to salt stress

Ubiquitination pathway is a way of protein modification after translation, which plays important roles in abiotic stress and hormone-singal sensing. E3 ligases confer specificity to ubiquitination by recognizing target substrates and mediating transfer of ubiquitin from an E2 ubiquitin conjugating enzyme to substrate. In this work, we studied the expression and function of AtTR1 (At3g47550), which shares 86% to its homologues in Brassica napus (BnTR1). In vitro self-ubiquitination assay demonstrated that the RINGv domain protein possess an E3 ligase activity. The analysis of expression pattern revealed that AtTR1 was dominantly accumulated in plant tissues and highly induced by salt (200mmol NaCl). The result indicated a specific role of AtTR1 in adaptation to salt stress. In addition, AtTR1 was cloned with PCR and constructed on the plant expression Vector pZH01. RT-PCR analysis of hygromycin resistant plants showed that AtTR1 gene had been integrated into the genome of attr1 mutant. These results provided important information for studying biological functions of AtTR1.