摘要
[目的]分析玉米ZmCIPK42序列,诱导蛋白表达,并获得纯化的蛋白。[方法]利用蛋白分析软件分析ZmCIPK42性质及磷酸化作用位点,构建ZmCIPK2-pET30a原核表达载体,用IPTG诱导蛋白表达,用NiNTA树脂纯化蛋白。[结果]ZmCIPK42具有典型的CIPK家族基因的N端激酶结构域和C端NAF调控域,激酶结构域内有较多磷酸化位点,原核表达的ZmCIPK42主要以包涵体形式存在,用Ni-NTA树脂可以纯化获得有活性的ZmCIPK42蛋白。[结论]28℃诱导后ZmCIPK42可以有活性的蛋白。用His作为标签纯化ZmCIPK42蛋白,可高效获得大量纯化蛋白。
[ Objective ] To analyze ZmCIPK42 sequence of maize, induce protein expression and obtain purified protein. [ Method ] Protein analysis software was used to analyze the properties and phosphorylation sites of ZmCIPK42, and to construct prokaryotic expression vector ZmCIPK42 -pET30a induced by IPTG, and then Ni - NTA resin was used for protein purification. [ Result] ZmCIPK42 as a typical CIPK family gene has N terminal kinase and C end NAF domain, and expression of ZmCIPK42 - His in DE3 shows that it is mainly in the form of inclusion. Using Ni - NTA resin can help us to get plenty of purified protein. [ Conclusion] Under 28℃, ZmCIPK42 can produce activity protein effectively. ＂His＂ was used as a protein purification tag, which can help us to obtain purified protein rapidly.
出处
《新疆农业科学》
CAS
CSCD
北大核心
2013年第1期33-37,共5页
Xinjiang Agricultural Sciences
基金
新疆维吾尔自治区自然科学基金(2009211B30)
新疆维吾尔自治区高技术研究计划项目(201011109)
