Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Transferase other than ribonuclease
Patent
1995-10-02
1999-08-17
Hendricks, Keith D.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Transferase other than ribonuclease
4352523, 43525233, 435325, 435419, 536 232, C12N 912, C12N 1554
Patent
active
059393017
ABSTRACT:
The invention relates to a substantially pure thermostable DNA polymerase from Thermotoga neapolitana (Tne) and mutants thereof. The Tne DNA polymerase has a molecular weight of about 100 kilodaltons and is more thermostable than Taq DNA polymerase. The mutant Tne DNA polymerase has at least one mutation selected from the group consisting of (1) a first mutation that substantially reduces or eliminates 3'.fwdarw.5' exonuclease activity of said DNA polymerase; (2) a second mutation that substantially reduces or eliminates 5'.fwdarw.3' exonuclease activity of said DNA polymerase; (3) a third mutation in the O helix of said DNA polymerase resulting in said DNA polymerase becoming non-discriminating against dideoxynucleotides. The present invention also relates to the cloning and expression of the wild type or mutant Tne DNA polymerase in E. coli, to DNA molecules containing the cloned gene, and to host cells which express said genes. The Tne DNA polymerase of the invention may be used in well-known DNA sequencing and amplification reactions.
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A copy
Chatterjee Deb K.
Hughes, Jr. A. John
Hendricks Keith D.
Life Technologies Inc.
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