Thermostable uracil DNA glycosylase and methods of use

Details Thermostable uracil DNA glycosylase and methods of use Thermostable uracil DNA glycosylase and methods of use

1997-09-09

1999-03-30

Patterson, Jr., Charles L.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

4353201, 43525233, 435476, 435488, 536 232, C12N 924, C12N 1556

Patent

active

058887954

ABSTRACT:
A novel uracil DNA glycosylase enzyme (referred to as Bpa UDG) has been identified in Bacillus pallidus and the gene encoding Bpa UDG has been cloned, sequenced and expressed to produce a recombinant UDG protein. The enzyme is thermostable and exhibits reaction kinetics similar to E. coli UDG. It is effectively inhibited by B. subtilis UGI.
Bpa UDG may be used to inactivate contaminating amplicons in nucleic acid amplification reactions, particularly at higher reaction temperatures. It may also be used to generate Bpa UDG-specific antibodies for purification of Bpa-UDG or for detecting Bpa UDG in a sample. Certain Bpa UDG antibodies may inactivate the enzyme and may therefore be useful as substitutes for UGI or heat, or in combination with UGI and/or heat, for controlling UDG activity in a reaction.

REFERENCES:
P. Glaser, et al. "Bacillus subtilis genome project: cloning and sequencing of the 97 kb region from 325.degree. to 333.degree." Molec. Microbiol. 10:371-384 (1993).
O. K. Kaboev, et al. "Uracil-DNA Glycosylase From Bacillus stearothermophilus" FEBS Lett. 132:337-340 (1981).
A. Koulis, et al. "Uracil-DNA glycosylase activities in hyperthermophilic micro-organisms" FEMS Microbiol. Lett. 143:267-271 (1996).
M. C. Longo, et al. "Use of uracil DNA glycosylase to control carry-over contamination in polymerase chain reactions" Gene 93:125-128 (1990).
U. Varshney, et al. "Sequence Analysis, Expression, and Conservation of Escherichia coli Uracil DNA Glycosylase and Its Gene (ung)" J. Biol. Chem. 263:7776-7784 (1988).
O. K. Kaboev, et al. "Uracil-DNA Glycosylase of Thermophilic Thermothrix thiopara" J. Bacteriol. 164:421-424 (1985).
R. D. Fleischmann, et al. "Whole-Genome Random Sequencing and Assembly of Haemophilus influenzae Rd" Science 269:496-512 (1995).
V. Mejean, et al. "Nucleotide sequence of the Streptococcus pneumoniae ung gene encoding uracil-DNA glycosylase" Nucl. Acids Res. 18:6693 (1990).
UCL Protein Crystallography Group "Uracil-DNA Glycosylase". http://bsmcha 1.biochem.ucl.ac.uk/bsm/xtal/udgase.html.
NCBI Complete Genomes (Haemophilus influenzae). www.ncbi.nim.nih.gov/cgi-bin/Complete Genomes/prot?HIN1155.

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