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The Large Fragment of Escherichia coli DNA Polymerase I Can Synthesize DNA Exclusively from Fluorescently Labeled Nucleotides
Author(s) -
Brakmann Susanne,
Nieckchen Petra
Publication year - 2001
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/1439-7633(20011001)2:10<773::aid-cbic773>3.0.co;2-s
Subject(s) - klenow fragment , dna polymerase i , dna polymerase , dna clamp , polymerase , nucleotide , escherichia coli , biology , dna polymerase ii , dna , microbiology and biotechnology , biochemistry , primer (cosmetics) , primase , chemistry , polymerase chain reaction , exonuclease , reverse transcriptase , gene , organic chemistry
A highly sensitive fluorescence‐based assay for screening DNA polymerase libraries (see schematic picture) revealed a surprising activity of the wild‐type Klenow fragment of Escherichia coli DNA polymerase I: This enzyme, which has emerged during natural evolution as a polymerase “fit” for natural substrates, retains full activity in the sole presence of artificial nucleotides that are labeled with a fluorescent dye of the rhodamine type. This unusual finding opens up the horizon for generating DNA exclusively from rhodamine‐labeled analogues—a necessary prerequisite to realizing the idea of “single‐molecule sequencing”.