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

doi:10.1007/s00792-004-0384-5

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 journal › Article › peer-review

28 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 language	English
Pages (from-to)	243-251
Number of pages	9
Journal	Extremophiles
Volume	8
Issue number	3
DOIs	https://doi.org/10.1007/s00792-004-0384-5
Publication status	Published - Jun 2004

Keywords

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

Access to Document

10.1007/s00792-004-0384-5

Cite this

@article{6ad9360da01a40a69f7874494a2c863c,

title = "Thermophilic bacterial DNA polymerases with reverse-transcriptase activity",

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.",

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

author = "H Shandilya and K Griffiths and EK Flynn and M Astatke and PJ Shih and JE Lee and GF Gerard and MD Gibbs and PL Bergquist",

year = "2004",

month = jun,

doi = "10.1007/s00792-004-0384-5",

language = "English",

volume = "8",

pages = "243--251",

journal = "Extremophiles",

issn = "1431-0651",

publisher = "Springer, Springer Nature",

number = "3",

}

TY - JOUR

T1 - Thermophilic bacterial DNA polymerases with reverse-transcriptase activity

AU - Shandilya, H

AU - Griffiths, K

AU - Flynn, EK

AU - Astatke, M

AU - Shih, PJ

AU - Lee, JE

AU - Gerard, GF

AU - Gibbs, MD

AU - Bergquist, PL

PY - 2004/6

Y1 - 2004/6

N2 - 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.

AB - 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.

KW - DNA polymerase

KW - reverse transcriptase

KW - thermophilic bacteria

KW - HUMAN-IMMUNODEFICIENCY

KW - ESCHERICHIA-COLI

KW - KLENOW FRAGMENT

KW - MESSENGER-RNA

KW - GENE-PRODUCT

KW - KRAFT PULP

KW - AMPLIFICATION

KW - PCR

U2 - 10.1007/s00792-004-0384-5

DO - 10.1007/s00792-004-0384-5

M3 - Article

C2 - 15197605

SN - 1431-0651

VL - 8

SP - 243

EP - 251

JO - Extremophiles

JF - Extremophiles

IS - 3

ER -

Thermophilic bacterial DNA polymerases with reverse-transcriptase activity

Abstract

Keywords

Access to Document

Fingerprint

Cite this