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Thermophilic bacterial DNA polymerases with reverse-transcriptase activity. 2004
Harini Shandilya, and
Kate Griffiths, and
Elizabeth K Flynn, and
Mekbib Astatke, and
Po-Jen Shih, and
Jun E Lee, and
Gary F Gerard, and
Moreland D Gibbs, and
Peter L Bergquist
Invitrogen Corporation, 1600 Faraday Avenue, Carlsbad, CA 92008, USA.
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.
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