DNA polymerase and Dna polymerase derived 3′-5′exonuclease
AssigneeUniversitetet I Tromsø—Norges Arktiske Universitet
InventorsAtle Noralf Larsen, Yvonne Piotrowski
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Abstract
The present invention relates to enzymes having DNA polymerase and 3′-5′ exonuclease activities. In particular, the present invention relates to a heat labile enzyme possessing a DNA polymerase II activity and a 3′-5′ exonuclease activity of marine origin. Furthermore, the present invention relates to a DNA polymerase primarily exerting a 3′-5′ activity, i.e. where the polymerase activity is absent. The present invention furthermore relates to the use of the exonuclease activity to degrade the 3′-5′ strand of double stranded DNA to perform single stranded overhang, e.g. in recombinant cloning processes, or in processes for removal of contaminating nucleic acid molecules.
Claims12
An isolated DNA polymerase derived 3′-5′exonuclease comprising an amino acid sequence which is at least 95% sequence identical over the entire length of the sequence with SEQ ID NO:
- 1, wherein
- the amino acid in position 442 is selected from the group consisting of Glu, Ala, Gly, Val, Leu, and Ile,
- the amino acid in position 445 is selected from the group consisting of Arg, Lys, and His,
- and/or
- the amino acid in position 568 is selected from the group consisting of Glu, Ala, Gly, Val, Leu, and Ile,
- wherein in the presence of dsDNA and dNTPs at 25° C., 5′ overhangs are produced in 5 to 30 minutes.
Dependencies are inferred from the claim text.