百脉根泛素结合酶LjE2的表达及活性鉴定
Expression and activity identification of ubiquitin binding enzyme LjE2 of Lotus japonicus

为了研究泛素化作用在豆科植物共生固氮过程中的调控功能和作用机制，利用拟南芥泛素结合酶E2的序列，在百脉根基因组上鉴定得到14个编码E2（LjE2）的基因，从LjE2中克隆到11个基因并构建了融合表达载体pET LjE2s:His。通过大肠杆菌表达系统，表达并纯化到10个LjE2:His融合蛋白。经体外泛素化及Western blot分析，在10个LjE2中有9个具有泛素结合酶活性，1个（LjE2 5）未检测到泛素结合酶活性。
Fourteen genes encoding E2(LjE2) in Lotus japonicus genome were identified based on the sequence of E2 in Arabidopsis to study the function and mechanism of ubiquitination in the process of symbiotic nitrogen fixation in legumes. Among them,eleven LjE2s were cloned to construct the fusion expression vectors pET LjE2s:His. Ten LjE2 proteins were successfully expressed and purified under the prokaryotic expression system. The results of ubiquitination in vitro and Western blotting showed that nine LjE2s had ubquitin conjugation enzyme activity and one had no detectable ubquitin conjugation enzyme activity.