中文摘要：

为了研究E3连接酶AtTR1和非生物胁迫应答的相关蛋白AtCPK28、AtCPK32之间关系，本文构建原核表达载体pET28a—AtCPK28和pET28a-AtCPK32，并在大肠杆菌中表达了AtTR1、AtCPK28和AtCPK32重组蛋白，利用亲和层析纯化相应融合蛋白．将AtCPK28和AtCPK32蛋白分别加入含有ATP、泛素、E1、E2和AtTR1的体外泛素反应体系中，western blot检测到AtCPK28和AtCPK32有多聚泛素化条带，说明AtTR1在体外能多泛素化修饰AtCPK28和AtCPK32．结果表明AtCPK28和AtCPK32可能是AtTR1调控植物抗逆信号中的下游靶蛋白．

英文摘要：

In order to analyze the relationship between E3 ligase AtTR1 and abiotic stress response related proteins AtCPK28 and AtCPK32,we prepared the prokaryotic expression plasmids pET28a-AtCPK28 and pET28a-AtCPK32, and produced the AtTR1, AtCPK28 and AtCPK32 recombinant proteins in Escherichia coli using RT-PCR and molecular approaches. The recombinant proteins was also purified by Affinity Chromatography. In the presence of ATP,E1, E2 and AtTR1, Poly ubiquitination band was observed in the presence o~ purified AtCPK28 or AtCPK32 by western blot. The results showed that the AtCPK28 and AtCPK32 were multi-ubiquitylated by AtTR1 in vitro. It indicated that the AtCPK28 and AtCPK32 are potential downstream targets of AtTR1 in the regulation of stress resistance signaling pathways.