Nucleic acid polymerase for single-molecule sequencing and mutant thereof

Abstract

Provided is a novel nucleic acid polymerase that can be used for single-molecule sequencing technology. The foregoing polymerase belongs to the Phi29 polymerase family, but the sequence homology thereof with other reported polymerases for sequencing technology is less than 60%. After characterization and identification, the polymerase shows good comprehensive performance in aspects such as thermal stability, extension capability at room temperature, extension processivity, ability to utilize modified nucleotide substrates / chain replacement function catalytic kinetics for same, and electrolyte resistance, and is suitable for single-molecule sequencing technology based on the "sequencing by synthesis" scheme. A beneficial mutant strain of the wild-type polymerase is further provided, which exhibits better kinetic characteristics and can increase sequencing accuracy and throughput.

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