Search results for "In vitro recombination"

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Generation of reporter plasmids containing defined base modifications in the DNA strand of choice

2012

Physiological effects of DNA bases other than A, G, C, and T as well as ways of removal of such bases from genomes are studied intensely. Methods for targeted insertion of modified bases into DNA, therefore, are highly demanded in the fields of DNA repair and epigenetics. This article describes efficient procedures for incorporation of modified DNA bases into a plasmid-borne enhanced green fluorescent protein (EGFP) gene. The procedure exploits excision of a stretch of 18 nt from either the transcribed or nontranscribed DNA strand with the help of the sequence-specific nicking endonucleases Nb.Bpu10I and Nt.Bpu10I. The excised single-stranded oligonucleotide is then swapped for a synthetic …

GuanineDNA clampBase pairGreen Fluorescent ProteinsOligonucleotidesBiophysicsDNACell BiologyNicking enzymeBiologyBiochemistryDNA GlycosylasesHeavy strandBiochemistryDNA glycosylaseCoding strandPrimer (molecular biology)UracilBase PairingMolecular BiologyIn vitro recombinationPlasmidsAnalytical Biochemistry

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