生物技术通报Issue(8)：88-93,6.DOI:10.13560/j.cnki.biotech.bull.1985.2015.08.013
拟南芥SPX1蛋白原核表达及纯化分析
Prokaryotic Expression and Purification of AtSPX1 Protein in Arabidopsis thaliana
摘要
Abstract
The proteins containing SPX domain exist widely in eukaryotes, and the functions of this kind of proteins are not clear yet, however some of them are involved in phosphorus signaling and some in iron signaling. SPX proteins inArabidopsis thalianacan be divided into 4 families. AtSPX1 used in this paper belongs to the family containing only SPX domain, while other 3 families containing extra gene sequences. Phylogenetic analysis showed that amino acid encoded by AtSPX1 was close with dicots in sequence, but distant from monocots. In order to reveal the relationship between structural property and biological function, we carried out solubility expression experiments of the proteinsin vitro, constructed prokaryotic expression vectorin vitro, and had the high soluble expression of the protein inEscherichia coli’scells. The expressed His-tag proteins allowed the purification more convenient, i.e, the inserted SUMO fusion protein tag could be digested by protease, and the target protein could be purified by ammonium sulfate precipitation. The further purification was completed using size exclusion chromatographic column, which yielded a profile corresponding to a monomeric AtSPX1 in solution. This work provided a strategy for the purification of AtSPX1 protein.关键词
SPX结构域/原核表达/蛋白纯化/拟南芥Key words
SPX domain/prokaryotic expression/protein purification/Arabidopsis thaliana引用本文复制引用
胡涛,安艳,吕群丹,徐英武..拟南芥SPX1蛋白原核表达及纯化分析[J].生物技术通报,2015,(8):88-93,6.基金项目
国家自然科学基金项目（31270715），浙江农林大学科研发展基金项目 （）
