Cold-shock induced high-yield protein production in Escherichia coli
Autor: | Thomas Acton, Tapas K. Mal, Mitsuhiko Ikura, G. V. T. Swapna, Ahmad Khorchid, Masanori Takayama, Haiping Ke, Sangita Phadtare, Bing Xia, Gaetano T. Montelione, Guoliang Qing, Li Chung Ma, Masayori Inouye |
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Rok vydání: | 2004 |
Předmět: |
Protein Conformation
Genetic Vectors Molecular Sequence Data Biomedical Engineering Bioengineering Biology medicine.disease_cause Applied Microbiology and Biotechnology Protein structure Bacterial Proteins Gene expression Escherichia coli Protein biosynthesis medicine Animals Humans Amino Acid Sequence Cloning Molecular Promoter Regions Genetic Nuclear Magnetic Resonance Biomolecular Peptide sequence Gene Cloning Expression vector Proteins Molecular biology Cold Temperature Biochemistry Isotope Labeling Protein Biosynthesis Molecular Medicine Biotechnology |
Zdroj: | Nature Biotechnology. 22:877-882 |
ISSN: | 1546-1696 1087-0156 |
Popis: | 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. |
Databáze: | OpenAIRE |
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