Abstract: Provided is a recombinant KOD polymerase. A KOD polymerase mutant is a protein as follows: the protein is a protein having DNA polymerase activity that is obtained by modifying amino acid residues in at least one of the following 48 positions in the amino acid sequence of KOD DNA polymerase GH78: 267, 326, 347, 349, 353, 375, 378, 379, 380, 385, 451, 452, 453, 454, 457, 461, 465, 470, 474, 477, 478, 479, 480, 482, 484, 485, 486, 493, 496, 497, 514, 574, 584, 605, 610, 630, 665, 666, 667, 674, 676, 680, 682, 698, 707, 718, 723 and 729, without changing other amino acid sequences; and compared with KOD DNA polymerase GH78, with regard to catalysis, the recombinant DNA polymerase exhibits a faster reaction rate, better catalytic efficiency, better affinity and other advantages, thereby improving the reaction rate of DNA polymerase in sequencing and increasing the reaction read length.

Abstract: Provided are a thermostable B-family DNA polymerase mutant and an application thereof. The thermostable B-family DNA polymerase mutant is provided to mutate amino acid residues in at least two of three sites of group A, i.e., 408, 409, and 410 in an amino acid sequence of thermostable B-family DNA polymerases to obtain proteins having DNA polymerase activity. A-group sites (408/409/410), B-group sites (451/485), C-group sites (389/383/384), D-group sites (589/67/680), E-group sites (491/493/494/497), and F-group sites (474/478/486/480/484) of several B-family DNA polymerases are analyzed to design the mutant.