Analysis and Suppression of DNA Polymerase Pauses Associated with a Trinucleotide Consensus

We have studied a novel class of DNA sequences that cause DNA polymerases to pause. These sequences have the central consensus Py-G-C and are not necessarily adjacent to hairpins in the DNA template. Since most consensus sequences do not cause pauses under standard conditions, additional template fe...

Full description

Saved in:
Bibliographic Details
Published in:Nucleic acids research 1996-07, Vol.24 (14), p.2774-2781
Main Authors: Mytelka, Daniel S., Chamberlin, Michael J.
Format: Article
Language:English
Subjects:
Bacteriophage T7 - enzymology
Base Sequence
Betaine - pharmacology
Consensus Sequence
DNA, Bacterial - chemistry
DNA, Bacterial - metabolism
DNA-Directed DNA Polymerase - drug effects
DNA-Directed DNA Polymerase - metabolism
Enzyme Stability
Molecular Sequence Data
Nucleic Acid Conformation
Nucleic Acid Synthesis Inhibitors
Sarcosine - pharmacology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We have studied a novel class of DNA sequences that cause DNA polymerases to pause. These sequences have the central consensus Py-G-C and are not necessarily adjacent to hairpins in the DNA template. Since most consensus sequences do not cause pauses under standard conditions, additional template features must exist that make it difficult to incorporate nucleotides at these positions. We believe that these pauses result from constraints that make the conformation change involved in nucleotide selection more difficult. These pauses can obscure parts of DNA sequencing ladders and prevent DNA amplification by the polymerase chain reaction. The addition of betaine, and some related compounds, relieves these pauses.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/24.14.2774