RECOMBINANT POLYMERASES WITH INCREASED PHOTOTOLERANCE

摘要

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

主权项

1. A composition comprising a φ29-type (phi29-type) recombinant DNA polymerase, which recombinant polymerase comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:2, and which recombinant polymerase comprises one or more mutation selected from the group consisting of an amino acid substitution at position S191, an amino acid substitution at position F436, an amino acid substitution at position T438, an amino acid substitution at position A444, an amino acid substitution at position S524, an L139R substitution, a D507Q substitution, and a T520H substitution, wherein identification of positions is relative to SEQ ID NO:2, and wherein said polymerase exhibits polymerase activity.