Cold-shock induced high-yield protein production in Escherichia coli

被引：277

作者：

Qing, GL

Ma, LC

Khorchid, A

Swapna, GVT

Mal, TK

Takayama, MM

Xia, B

Phadtare, S

Ke, HP

Acton, T

Montelione, GT

Ikura, M

Inouye, M

机构：

[1] Univ Med & Dent New Jersey, Robert Wood Johnson Med Sch, Dept Biochem, Piscataway, NJ 08854 USA

[2] Rutgers State Univ, NE Struct Genom Consortium, Piscataway, NJ 08854 USA

[3] Rutgers State Univ, Ctr Adv Biotechnol & Med, Dept Mol Biol & Biochem, Piscataway, NJ 08854 USA

[4] Univ Toronto, Ontario Canc Inst, Div Mol & Struct Biol, Toronto, ON M5G 2M9, Canada

[5] Univ Toronto, Dept Med Biophys, Toronto, ON M5G 2M9, Canada

[6] Takara Bio Inc, Shiga 5202193, Japan

来源：

NATURE BIOTECHNOLOGY | 2004年 / 22卷 / 07期

关键词：

D O I：

10.1038/nbt984

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Overexpression of proteins in Escherichia coli at low temperature improves their solubility and stability(1,2). Here, we apply the unique features of the cspA gene to develop a series of expression vectors, termed pCold vectors, that drive the high expression of cloned genes upon induction by cold-shock. Several proteins were produced with very high yields, including E. coli EnvZ ATP-binding domain (EnvZ-B) and Xenopus laevis calmodulin (CaM). The pCold vector system can also be used to selectively enrich target proteins with isotopes to study their properties in cell lysates using NMR spectroscopy. We have cloned 38 genes from a range of prokaryotic and eukaryotic organisms into both pCold and pET14 (ref. 3) systems, and found that pCold vectors are highly complementary to the widely used pET vectors.

引用

页码：877 / 882

页数：6

共 18 条

[1] Selectively labeling the heterologous protein in Escherichia coli for NMR studies:: A strategy to speed up NMR spectroscopy [J].