Novel DNA-Ploymerases

摘要

The technology provided herein relates to novel variants of DNA-Polymerases exhibiting high termo-stability as well as a strong strand displacement activity; to nucleic acid molecules encoding said DNA-Polymerases, vectors, host cells containing the nucleic acids and methods for preparation and producing such enzymes; compositions comprising at least one of the DNA-Polymerases; and methods for using such enzymes in DNA sequencing and/or DNA amplification processes.

主权项

1. A modified DNA-polymerase within the Type-A family of polymerases, wherein the polymerase comprises substitutions of at least two amino acid residues in the amino acid sequence of the naturally occurring DNA-polymerase at positions corresponding to 738 and 743 relative to the numbering of the amino acid sequence of wild-type Thermus aquaticus (Taq) DNA-polymerase (SEQ ID NO: 1), wherein the substitutions are 738F and 743F, said modified DNA polymerase, or a homolog thereof, has an increased strand display activity relative to the corresponding naturally occurring unmodified DNA-polymerase, said DNA modified polymerase, or a homolog thereof, is thermostable at a temperature of at least 80° C. and/or has a reduced 5′-3′ exonuclease activity, and the naturally occurring Type-A DNA polymerase is a thermostable Thermus species DNA polymerase.