Cold-shock induced high-yield protein production in Escherichia coli
Top Cited Papers
- 13 June 2004
- journal article
- letter
- Published by Springer Nature in Nature Biotechnology
- Vol. 22 (7) , 877-882
- https://doi.org/10.1038/nbt984
Abstract
Overexpression of proteins in Escherichia coli at low temperature improves their solubility and stability1,2. 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.Keywords
This publication has 14 references indexed in Scilit:
- Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genesPublished by Elsevier ,2004
- Identification and Biosynthesis of Cyclic Enterobacterial Common Antigen in Escherichia coliJournal of Bacteriology, 2003
- Nonsense Mutations in cspA Cause Ribosome Trapping Leading to Complete Growth Inhibition and Cell Death at Low Temperature in Escherichia coliJournal of Biological Chemistry, 2001
- Selectively Labeling the Heterologous Protein in Escherichia coli for NMR Studies: A Strategy to Speed Up NMR SpectroscopyJournal of Magnetic Resonance, 2001
- High-level production of uniformly 15N-and 13C-enriched fusion proteins in Escherichia coliJournal of Biomolecular NMR, 1996
- Rapid screening for structural integrity of expressed proteins by heteronuclear NMR spectroscopyProtein Science, 1996
- NMRPipe: A multidimensional spectral processing system based on UNIX pipesJournal of Biomolecular NMR, 1995
- Calcium-induced conformational transition revealed by the solution structure of apo calmodulinNature Structural & Molecular Biology, 1995
- The 13C Chemical-Shift Index: A simple method for the identification of protein secondary structure using 13C chemical-shift dataJournal of Biomolecular NMR, 1994
- Low temperature cultivation of Escherichia coli carrying a rice lipoxygenase L‐2 cDNA produces a soluble and active enzyme at a high levelFEBS Letters, 1990