|Nature Biotechnology (2004) 22(7):877-82|
|Northeast Structural Genomics Consortium|
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Overexpression of proteins in Escherichia coli at low temperature improves their solubility and stability. ...
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.
|chemistry genetics metabolism |
|Escherichia coli Genetic Vectors Humans Molecular Sequence Data Promoter Regions, Genetic Protein Conformation Cloning, Molecular Isotope Labeling Animals Amino Acid Sequence Proteins Nuclear Magnetic Resonance, Biomolecular Bacterial Proteins Cold Temperature Protein Biosynthesis |
|199 (Last update: 05/27/2017 11:38:17am)|
|Nat Biotechnol. 2004 Jul;22(7):877-82. Epub 2004 Jun 13.|