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Overexpression, purification and preliminary X-ray diffraction analysis of the controller protein C. Csp231I from Citrobacter sp. RFL231.

Acta Crystallogr Sect F Struct Biol Cryst Commun.. 2009-09;  65(9):898-91
Streeter SD, McGeehan JE, Kneale GG. Biophysics Laboratories, Institute of Biomedical and Biomolecular Sciences, School of Biological Sciences, University of Portsmouth, Portsmouth PO1 2DY, England.
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摘要

Restriction–modification controller proteins play an essential role in regulating the temporal expression of restriction–modification genes. The controller protein C.Csp231I represents a new class of controller proteins. The gene was sublconed to allow overexpression in Escherichia coli. The protein was purified to homogeneity and crystallized using the hanging-drop vapour-diffusion method. The crystals diffracted to 2.0 Å resolution and belonged to space group P21. An electrophoretic mobility-shift assay provided evidence of strong binding of C.Csp231I to a sequence located upstream of the csp231IC start codon.

关键词

DNA-binding proteins; restriction-modification systems; transcription; gene regulation.
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