Nature Biotechnology
22, 877 - 882 (2004)
Published online: 13 June 2004; | doi:10.1038/nbt984
Cold-shock induced high-yield protein production in Escherichia coliGuoliang Qing1, Li-Chung Ma2, 3, Ahmad Khorchid4, G V T Swapna2, 3, Tapas K Mal4, Masanori Mitta Takayama5, Bing Xia1, Sangita Phadtare1, Haiping Ke1, Thomas Acton2, 3, Gaetano T Montelione1, 2, 3, Mitsuhiko Ikura4
& Masayori Inouye1, 21
Department of Biochemistry, Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, New Jersey 08854, USA. 2
Northeast Structural Genomics Consortium, Rutgers University, 679 Hoes Lane, Piscataway, New Jersey 08854, USA. 3
Department of Molecular Biology and Biochemistry, Center for Advanced Biotechnology and Medicine (CABM), Rutgers University, 679 Hoes Lane, Piscataway, New Jersey 08854, USA. 4
Division of Molecular and Structural Biology, Ontario Cancer Institute and Department of Medical Biophysics, University of Toronto, 610 University Avenue, Toronto, Ontario M5G 2M9, Canada. 5
Takara Bio Inc., Otsu, Shiga, 520-2193, Japan.
Correspondence should be addressed to Masayori Inouye inouye@umdnj.eduOverexpression 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.
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