Patent ID: 6228628
Filing Date: 2001-05-08
Classification: C07K,C12N,C12Q

Abstract:
A thermostable DNA polymerase consisting of an N-terminal region and a C-terminal region,wherein said N-terminal region consists of amino acids 1 through n of a Thermus species DNA polymerase, wherein n is an amino acid corresponding to an amino acid m of a Thermatoga maritima (Tma) DNA polymerase described by SEQ ID NO: 10, wherein m is between 137 and 291;wherein said C-terminal region consists of amino acids m+1 through 893 of said Tma DNA polymerase;wherein said N-terminal region is modified by at least one point mutation, each characterized in that it substantially reduces or eliminates 5'-nuclease activity when present in said Thermus species DNA polymerase, or said C-terminal region is modified by at least one point mutation within the region that is amino acids m+1 to 291 of Tma DNA polymerase, each characterized in that it substantially reduces or eliminates 5'-nuclease activity when present in Tma DNA polymerase;wherein said C-terminal region is modified by at least one point mutation, each characterized in that it substantially reduces 3' to 5' exonuclease activity when present in Tma DNA polymerase; andwherein said C-terminal region is modified to contain a tyrosine at amino acid 730.