Cloning and expression of T5 DNA polymerase reduced in 3'-to-5'

Details Cloning and expression of T5 DNA polymerase reduced in 3'-to-5' Cloning and expression of T5 DNA polymerase reduced in 3'-to-5'

1990-03-16

1996-07-30

Knode, Marian C.

Chemistry: molecular biology and microbiology

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

Transferase other than ribonuclease

435193, 4351723, C12N 1500, C12N 912

Patent

active

055410995

ABSTRACT:
The present invention discloses a recombinant DNA molecule having a structural gene encoding a processive, thioredoxin-independent DNA polymerase that is substantially reduced in processive 3'-to-5' DNA exonuclease activity, a promoter, and an origin of replication. A method for producing this enzyme is also disclosed, as is the protein produced by this process. This invention is exemplified by expression of exonuclease.sup.- T5 DNA polymerase in E. coli.

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patent: 4795699 (1989-01-01), Tabor et al.
patent: 4889818 (1989-12-01), Gelfand et al.
Boehringer Mannheim, p. 317.
Bernas et al., "A Conserved 3'-5' Exonuclease Active Sets in Prokaryotic and Eukaryotic DNA Polymerase", Cell, vol. 59, pp. 219-228, 1989.
Das et al., "Mechanism of Primer-Template-dependent Conversion of d NTP & d NMP by T5 DNA Polymerase", J. of Biol Chem, vol. 255, No. 15 pp. 7149-7154 1980.
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Tabor et al., Selective Inactivation of the Exonuclease Activity of Bacteriophage T7 DNA Polymerase by in Vitro Mutagenesis, The Journal of Biological Chemistry, 264(11)6447-6458 (1989).

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