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

Guoliang Qing, Li Chung Ma, Ahmad Khorchid, G. V.T. Swapna, Tapas Mal, Masanori Mitta Takayama, Bing Xia, Sangita Phadtare, Haiping Ke, Thomas Acton, Gaetano T. Montelione, Mitsuhiko Ikura, Masayori Inouye

Research output: Contribution to journalArticle

221 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)877-882
Number of pages6
JournalNature Biotechnology
Volume22
Issue number7
DOIs
StatePublished - Jul 1 2004

Fingerprint

Escherichia coli
Shock
Proteins
Escherichia coli Proteins
Xenopus laevis
Calmodulin
Isotopes
Solubility
Genes
Magnetic Resonance Spectroscopy
Adenosine Triphosphate
Gene Expression
Temperature
Adenosinetriphosphate
Nuclear magnetic resonance spectroscopy

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

Cite this

Qing, G., Ma, L. C., Khorchid, A., Swapna, G. V. T., Mal, T., Takayama, M. M., ... Inouye, M. (2004). Cold-shock induced high-yield protein production in Escherichia coli. Nature Biotechnology, 22(7), 877-882. https://doi.org/10.1038/nbt984
Qing, Guoliang ; Ma, Li Chung ; Khorchid, Ahmad ; Swapna, G. V.T. ; Mal, Tapas ; Takayama, Masanori Mitta ; Xia, Bing ; Phadtare, Sangita ; Ke, Haiping ; Acton, Thomas ; Montelione, Gaetano T. ; Ikura, Mitsuhiko ; Inouye, Masayori. / Cold-shock induced high-yield protein production in Escherichia coli. In: Nature Biotechnology. 2004 ; Vol. 22, No. 7. pp. 877-882.
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Qing, G, Ma, LC, Khorchid, A, Swapna, GVT, Mal, T, Takayama, MM, Xia, B, Phadtare, S, Ke, H, Acton, T, Montelione, GT, Ikura, M & Inouye, M 2004, 'Cold-shock induced high-yield protein production in Escherichia coli', Nature Biotechnology, vol. 22, no. 7, pp. 877-882. https://doi.org/10.1038/nbt984

Cold-shock induced high-yield protein production in Escherichia coli. / Qing, Guoliang; Ma, Li Chung; Khorchid, Ahmad; Swapna, G. V.T.; Mal, Tapas; Takayama, Masanori Mitta; Xia, Bing; Phadtare, Sangita; Ke, Haiping; Acton, Thomas; Montelione, Gaetano T.; Ikura, Mitsuhiko; Inouye, Masayori.

In: Nature Biotechnology, Vol. 22, No. 7, 01.07.2004, p. 877-882.

Research output: Contribution to journalArticle

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Qing G, Ma LC, Khorchid A, Swapna GVT, Mal T, Takayama MM et al. Cold-shock induced high-yield protein production in Escherichia coli. Nature Biotechnology. 2004 Jul 1;22(7):877-882. https://doi.org/10.1038/nbt984