Highly efficient expression and purification system of small-size protein domains in Escherichia coli for biochemical characterization

	Highly efficient expression and purification system of small-size protein domains in Escherichia coli for biochemical characterization
	Bao, WJ; Gao, YG; Chang, YG; Zhang, TY; Lin, XJ; Yan, XZ; Hu, HY
刊名	PROTEIN EXPRESSION AND PURIFICATION
	2006
卷号	47 期号:2 页码:599-606
关键词	domain expression purification GB1 fusion protein biochemical characterization
通讯作者	Hu, HY (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Key Lab Proteom, Shanghai 200031, Peoples R China.,hyhu@sibs.ac.cn
英文摘要	It is often essential to focus the study on the small-size domains of large proteins in eukaryotic cells in the post-genomic era, but the low expression level, insolubility, and instability of the domains have been continuing to hinder the massive purification of domain peptides for structural and biological investigation. In this work, a highly efficient expression and purification system based on a small-size fusion partner GB I and histidine tag was utilized to solve these problems. Two vectors, namely pGBTNH and pGBH, were constructed to improve expression and facilitate purification. The linker and thrombin cleavage site have been optimized for minimal degradation during purification process. This system has been tested for eight domain peptides varying in size, linker, hydrophobicity, and predicted secondary structure. The results indicate that this system is achievable to produce these domain peptides with high solubility and stability for further biochemical characterization. Moreover, the fusion protein without the linker and thrombin cleavage site is also suitable for spectroscopic studies especially for NMR structural elucidation, if the target peptide is prone to precipitation or easily degraded during purification. This system will be beneficial to the research field of structure and function of small domain and peptide fragment. (c) 2005 Elsevier Inc. All rights reserved.
学科主题	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
类目[WOS]	Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
关键词[WOS]	DNA
收录类别	SCI
语种	英语
WOS记录号	WOS:000238277000032
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1666]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Bao, WJ,Gao, YG,Chang, YG,et al. Highly efficient expression and purification system of small-size protein domains in Escherichia coli for biochemical characterization[J]. PROTEIN EXPRESSION AND PURIFICATION,2006,47(2):599-606.
APA	Bao, WJ.,Gao, YG.,Chang, YG.,Zhang, TY.,Lin, XJ.,...&Hu, HY.(2006).Highly efficient expression and purification system of small-size protein domains in Escherichia coli for biochemical characterization.PROTEIN EXPRESSION AND PURIFICATION,47(2),599-606.
MLA	Bao, WJ,et al."Highly efficient expression and purification system of small-size protein domains in Escherichia coli for biochemical characterization".PROTEIN EXPRESSION AND PURIFICATION 47.2(2006):599-606.