| 玉米ZmCIPK42原核表达载体构建及蛋白纯化 |
| |
| 引用本文： | 陈勋基,陈果,黄全生.玉米ZmCIPK42原核表达载体构建及蛋白纯化[J].新疆农业科学,2013(1):33-37. |
| |
| 作者姓名： | 陈勋基 陈果 黄全生 |
| |
| 作者单位： | 新疆农业科学院核技术生物技术研究所 |
| |
| 基金项目： | 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) |
| |
| 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。
|
| 关 键 词： | 玉米 CIPK 蛋白纯化 |
Maize ZmCIPK42 Prokaryotic Expression Vector Construction and Its Protein Purification |
| |
| Affiliation: | CHEN Xun -ji,CHEN Guo,HUANG Quan -sheng (Research Institute of Nuclear and Biological Technologies,Xinjiang Academy of Agricultural Sciences, Urumqi 830091,China) |
| |
| Abstract: | 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-pET30αinduced 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 ZmCIPKAl - 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. |
| |
| Keywords: | maize CIPK protein purification |
| 本文献已被 CNKI 等数据库收录！ |
|
