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Protein Expr Purif. 1998 Feb;12(1):25-8.

An expression system of rat calmodulin using T7 phage promoter in Escherichia coli.

Author information

1
Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi, 470-11, Japan. nhayashi@fujita-hu.ac.jp

Abstract

An efficient expression system of rat calmodulin in Escherichia coli is presented. To express rat calmodulin cDNA, we employed a pET expression vector which contains the T7 phage promoter and terminator. After transformation of E. coli BL21(DE3) strain which carries T7 phage RNA polymerase inducible with isopropyl-beta-D-thiogalactopyranoside, induction of the expression, and chromatography of soluble proteins on a phenyl-Sepharose column, about 250 mg of recombinant rat calmodulin was obtained from 1 liter of E. coli culture. The recombinant calmodulin lacked the N-terminal methionine, and posttranslational modifications such as Nalpha-acetylation and methylation. This system facilitates the large amount preparation of calmodulin and the mutant proteins required for the structural analysis by NMR spectrometry and/or X-ray crystallography.

PMID:
9473453
DOI:
10.1006/prep.1997.0807
[Indexed for MEDLINE]

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