Thermophilic bacterial DNA polymerases with reverse-transcriptase activity

H Shandilya, K Griffiths, EK Flynn, M Astatke, PJ Shih, JE Lee, GF Gerard, MD Gibbs, PL Bergquist*

*Corresponding author for this work

    Research output: Contribution to journalArticle

    23 Citations (Scopus)

    Abstract

    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.

    Original languageEnglish
    Pages (from-to)243-251
    Number of pages9
    JournalExtremophiles
    Volume8
    Issue number3
    DOIs
    Publication statusPublished - Jun 2004

    Keywords

    • DNA polymerase
    • reverse transcriptase
    • thermophilic bacteria
    • HUMAN-IMMUNODEFICIENCY
    • ESCHERICHIA-COLI
    • KLENOW FRAGMENT
    • MESSENGER-RNA
    • GENE-PRODUCT
    • KRAFT PULP
    • AMPLIFICATION
    • PCR

    Cite this

    Shandilya, H., Griffiths, K., Flynn, EK., Astatke, M., Shih, PJ., Lee, JE., ... Bergquist, PL. (2004). Thermophilic bacterial DNA polymerases with reverse-transcriptase activity. Extremophiles, 8(3), 243-251. https://doi.org/10.1007/s00792-004-0384-5