IP Library › Granted Patent US 12,385,021
Granted Patent B2
US 12,385,021 · App. 16/957,276 · Granted Aug 12, 2025

Heat-resistant DNA polymerase mutant with high amplification activity

Inventors: Xiwen Jiang (Guangdong, CN); Aishan Liu (Guangdong, CN)
Assignee: DAAN GENE CO., LTD.
C12N9/1252C12Y207/07007
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Quick Facts
Patent No.
US 12,385,021
App. No.
16/957,276
Granted
Aug 12, 2025
Kind
B2
Abstract

The present invention provides a heat-resistant DNA polymerase mutant with high amplification activity. Particularly, the present invention uses protein directed evolution technology to construct a random mutation library for the polymerase active domain of Taq enzyme, and gradually adds screening pressure, so that unsuitable mutations will be eliminated naturally, and mutations with dominant traits will gradually accumulate. Finally, a series of amino acid sites and their mutations that are critical to Taq enzyme amplification and polymerization performance will be selected, and a Taq enzyme mutant with high amplification activity will be obtained.

Claims (9)

1. A mutated DNA polymerase,

wherein the mutated DNA polymerase is mutated based on the wild-type DNA polymerase as set forth in SEQ ID NO: 2,

wherein the amino acid sequence of the mutated DNA polymerase has at least 98% homology to SEQ ID NO: 2, and the mutated DNA polymerase comprises the mutations:

F495R and F749T (Mutant No. 12).

2. A method for preparing the mutated DNA polymerase of claim 1 , wherein the method comprises the steps of:

(i) culturing a host cell under suitable conditions to express the mutant DNA polymerase; and

(ii) isolating the mutated DNA polymerase,

wherein the host cell contains a chromosome integrated with the nucleic acid molecule encoding the mutated DNA polymerase of claim 1 .

3. A kit, wherein the kit comprises the mutated DNA polymerase of claim 1 .

Assignments (2)
CHANGE OF NAME Recorded Aug 6, 2021
From: DAAN GENE CO., LTD. OF SUN YAT-SEN UNIVERSITY
To: DAAN GENE CO., LTD.
Reel/Frame 057115/0835 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 23, 2020
From: JIANG, XIWEN; LIU, AISHAN
To: DAAN GENE CO., LTD. OF SUN YAT-SEN UNIVERSITY
Reel/Frame 053015/0435 →
Continuity (1)
Related Publication 20220325259A1 · Oct 13, 2022
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