منابع مشابه
DNA polymerase β : effects of gapped DNA substrates on dNTP specificity, fidelity, processivity and conformational changes1
Pre-steady-state kinetic analysis was used to compare the catalytic properties of DNA polymerase β (Pol β) for single-base gapfilling and regular duplex DNA synthesis. The rate of polymerization (k pol ) and the apparent equilibrium dissociation constant of dNTP (K d ) were determined with single-nucleotide gapped DNA substrates for all four possible correct base pairs and twelve possible incor...
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We have partially purified a DNA methyltransferase from human placenta using a novel substrate for a highly sensitive assay of methylation of hemimethylated DNA. This substrate was prepared by extensive nick translation of bacteriophage XP12 DNA, which normally has virtually all of its cytosine residues replaced by 5-methylcytosine (m5C). Micrococcus luteus DNA was just as good a substrate if i...
متن کاملDNA sequencing using polymerase substrate-binding kinetics
Next-generation sequencing (NGS) has transformed genomic research by decreasing the cost of sequencing. However, whole-genome sequencing is still costly and complex for diagnostics purposes. In the clinical space, targeted sequencing has the advantage of allowing researchers to focus on specific genes of interest. Routine clinical use of targeted NGS mandates inexpensive instruments, fast turna...
متن کاملSubstrate Rescue of DNA Polymerase β Containing a Catastrophic L22P Mutation
DNA polymerase (pol) β is a multidomain enzyme with two enzymatic activities that plays a central role in the overlapping base excision repair and single-strand break repair pathways. The high frequency of pol β variants identified in tumor-derived tissues suggests a possible role in the progression of cancer, making the determination of the functional consequences of these variants of interest...
متن کاملThe Mutational Specificity of DNA Polymerase - @ during in Vitro DNA
The frequency and specificity of mutations produced in uitro by eucaryotic DNA polymerase-@ have been determined in a forward mutation assay using a 250base target sequence in M13mp2 DNA. Homogeneous DNA polymerase-@, isolated from four different sources, produces mutations at a frequency of 4-6%/ single round of gap-filling DNA synthesis. DNA sequence analyses of 460 independent mutants res...
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ژورنال
عنوان ژورنال: Journal of Biological Chemistry
سال: 2009
ISSN: 0021-9258
DOI: 10.1074/jbc.m109.029843