Mutant Taq DNA polymerase and application thereof

Through the analysis of Taq enzyme and substrate binding sites and the AlphaFold3 prediction model, mutation sites such as D785G and R573K were screened out, and the mutant Taq DNA polymerase was constructed, which solved the problem of low amplification activity and efficiency of existing Taq enzymes when facing inhibitors and high GC content templates, achieving higher enzyme activity and a wider range of applications.

CN119709688AActive Publication Date: 2025-03-28HANGZHOU HUIXIN BIOTECHNOLOGY CO LTD +1
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Patent Information

Application Number
CN202510224433.3
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-02-27
Publication Date
2025-03-28
Estimated Expiration
2045-02-27

AI Technical Summary

Technical Problem

Existing Taq DNA polymerases have low amplification activity and efficiency when facing inhibitors and high GC content templates, resulting in poor results in some applications.

Method used

Through the analysis of Taq enzyme binding sites and substrate binding sites, combined with the AlphaFold3 prediction model, mutation sites such as D785G and R573K were screened to construct mutant Taq DNA polymerase to improve its tolerance to inhibitors and amplification ability of high GC content templates.

Benefits of technology

The enzyme activity of mutant Taq DNA polymerase can more effectively amplify the genome with high GC content, enhance the tolerance to inhibitors, and expand its application range in PCR reactions.

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Abstract

The invention discloses mutant Taq DNA polymerase and application thereof, and belongs to the technical field of biology. On one hand, the invention provides a mutant Taq DNA polymerase, and compared with the amino acid sequence of a wild Taq DNA polymerase, the mutant Taq DNA polymerase comprises a mutation site D785 and / or a mutation site R573; on the other hand, the invention provides application of the mutant Taq DNA polymerase in preparation of a PCR reaction reagent. The Taq DNA polymerase mutant disclosed by the invention is a mutant obtained by expression verification after the activity of a key mutation site is predicted by AlphaFold3. The double mutant strain obtained by the invention provides better Taq enzyme selection for genome amplification.
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