一种DNA聚合酶
By introducing amino acid substitution mutations at specific positions on DNA polymerase, the problems of decreased reaction rate and increased mismatch rate caused by Mn2+ ions were solved, enabling efficient identification and utilization of non-natural nucleotides under Mg2+ ion conditions, thus improving the accuracy and efficiency of high-throughput sequencing.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- CYGNUS BIOSCI BEIJING CO LTD
- Filing Date
- 2023-03-29
- Publication Date
- 2026-07-17
AI Technical Summary
Existing DNA polymerases are prone to oxidation and precipitation when using Mn2+ ions as cofactors, leading to a decrease in reaction rate, enhanced background fluorescence, and increased mismatch rate, which affects the accuracy and efficiency of high-throughput sequencing.
DNA polymerases are modified to recognize and utilize terminal phosphorylated fluorescently labeled non-natural nucleotides under Mg2+ ion conditions by introducing amino acid substitution mutations at specific positions, such as functionally or positionally equivalent substitutions at Lys464, Gln483, Glu578, or Glu580.
This technology enables efficient identification and utilization of non-natural nucleotides in the presence of Mg2+ ions, improving the accuracy and efficiency of sequencing reactions and expanding the application scope of fluorescence generation sequencing.
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Figure CN116286720B_ABST