Thermophilic bacterial DNA polymerases with reverse-transcriptase activity

Conserved motifs found in known bacterial polI DNA polymerase sequences were identified, and degenerate PCR primers were designed for PCR amplification of an internal portion of polI genes from all bacterial divisions. We describe here a method that has allowed the rapid identification and isolation...

Full description

Saved in:
Bibliographic Details
Published in:Extremophiles : life under extreme conditions 2004-06, Vol.8 (3), p.243-251
Main Authors: SHANDILYA, Harini, GRIFFITHS, Kate, FLYNN, Elizabeth K, ASTATKE, Mekbib, SHIH, Po-Jen, LEE, Jun E, GERARD, Gary F, GIBBS, Moreland D, BERGQUIST, Peter L
Format: Article
Language:English
Subjects:
Bacillus - enzymology
Bacillus - genetics
Bacillus caldolyticus
Bacteria
Bacteria - enzymology
Bacteria - genetics
Bacteriology
Base Sequence
Biochemistry
Biological and medical sciences
Cloning, Molecular
Clostridium - enzymology
Clostridium - genetics
Clostridium stercorarium
Deoxyribonucleic acid
DNA
DNA Polymerase I - genetics
DNA Polymerase I - metabolism
DNA Primers - genetics
DNA, Bacterial - genetics
Enzyme Stability
Fundamental and applied biological sciences. Psychology
Genes, Bacterial
Hot Temperature
Kinetics
Magnesium - pharmacology
Microbiology
Miscellaneous
Molecular Sequence Data
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
RNA-Directed DNA Polymerase - genetics
RNA-Directed DNA Polymerase - metabolism
Sequence Homology, Nucleic Acid
Space life sciences
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Conserved motifs found in known bacterial polI DNA polymerase sequences were identified, and degenerate PCR primers were designed for PCR amplification of an internal portion of polI genes from all bacterial divisions. We describe here a method that has allowed the rapid identification and isolation of 13 polI genes from a diverse selection of thermophilic bacteria and report on the biochemical characteristics of nine of the purified recombinant enzymes. Several enzymes showed significant reverse-transcriptase activity in the presence of Mg2+, particularly the polymerases from Bacillus caldolyticus EA1, Caldibacillus cellovorans CompA.2, and Clostridium stercorarium.
ISSN:1431-0651
1433-4909
DOI:10.1007/s00792-004-0384-5