Arabidopsis Calmodulin Expression Vectors

Calmodulin (CaM) is easily expressed and purified from cultures of transformed E. coli. The maps below illustrate two basic plasmids we constructed for expressing unfused CaM proteins. pETCaM2 is an early version that is constructed in pET5a and confers ampicillin resistance to transformed bacteria; pETCaM8 is a later version that is constructed in pET24d and confers kanamycin resistance to bacteria.

Vectors based on pET5a are available for expressing CaM2 (same as CaM3 and 5), CaM4 (same as CaM1) and CaM6. Vectors based on pET24d are available for expressing CaM7, CaM8 and CaM9. Sequences of the clones can be obtained as plain text files by clicking on the links. If you wish to obtain an expression clone, please read about the Uniform Biolgical Material Transfer Agreement (signed by the University of Illinois on March 11, 1998); the required paperwork and the transfer can be done using the Simple Letter Agreement.

Bacterial expression vectors for GFP-CaM2, GFP-CaM8 and GFP-CaM9 fusion proteins based on pET24d and GST-CaM2, GST-CaM8 and GST-CaM9 are also available. The GFP-CaM fusion proteins can be purified using the same protocol described for unfused CaM proteins. GST-CaM fusions have been purified using glutathione agarose affinity chromatography. The design of these vectors is shown in the figures below.

Our protocol for purifying recombinant CaM from E. coli can be downloaded through this link.

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