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

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...

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Detalles Bibliográficos
Autores principales: Streeter, S. D., McGeehan, J. E., Kneale, G. G.
Formato: Texto
Lenguaje:English
Publicado: International Union of Crystallography 2009
Materias:
Crystallization Communications
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2795595/
https://www.ncbi.nlm.nih.gov/pubmed/19724127
http://dx.doi.org/10.1107/S1744309109028681
Descripción
Sumario: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 P2(1). An electrophoretic mobility-shift assay provided evidence of strong binding of C.Csp231I to a sequence located upstream of the csp231IC start codon.

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