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Fusion of GFP to the M.EcoKI DNA methyltransferase produces a new probe of Type I DNA restriction and modification enzymes.

Abstract：

:We describe the fusion of enhanced green fluorescent protein to the C-terminus of the HsdS DNA sequence-specificity subunit of the Type I DNA modification methyltransferase M.EcoKI. The fusion expresses well in vivo and assembles with the two HsdM modification subunits. The fusion protein functions as a sequence-specific DNA methyltransferase protecting DNA against digestion by the EcoKI restriction endonuclease. The purified enzyme shows Förster resonance energy transfer to fluorescently-labelled DNA duplexes containing the target sequence and to fluorescently-labelled ocr protein, a DNA mimic that binds to the M.EcoKI enzyme. Distances determined from the energy transfer experiments corroborate the structural model of M.EcoKI.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Chen K,Roberts GA,Stephanou AS,Cooper LP,White JH,Dryden DT

doi

10.1016/j.bbrc.2010.06.069

subject

Has Abstract

pub_date

2010-07-23 00:00:00

pages

254-9

issue

2

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(10)01195-2

journal_volume

398

pub_type

杂志文章

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