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

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

Research output: Contribution to journalArticlepeer-review

297 Scopus citations


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
Issue number7
StatePublished - Jul 2004

All Science Journal Classification (ASJC) codes

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


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