DNA polymerase possessing continuous catalytic capacity and salt tolerance
US10047349B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Sep 5, 2016 |
| Grant date | Aug 14, 2018 |
| Priority date | — |
| Expiry date | Sep 5, 2036 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Y207/07007
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A DNA polymerase, having an amino acid sequence represented by SEQ ID No. 2, or a derivative of the amino acid sequence by substitution, deletion, or addition of at least one amino acid residue. The DNA polymerase is a hybrid DNA polymerase prepared by inserting a thioredoxin binding domain (TBD) of bacteriophage T7 DNA polymerase into a DNA polymerase I (Sau) of Staphylococcus aureus. A method for preparing the DNA polymerase includes: 1) determining a corresponding position and a target substitution sequence in Sau protein for the TBD of the bacteriophage T7 DNA polymerase; 2) devising and synthesizing a primer according to a gene sequence of Sau and a sequence TBD published by GenBank; 3) cloning the Sau-TBD segment acquired in (2) to an expression vector pTrc99A to construct a recombinant vector pTrc99A-Sau-TBD; and 4) transforming Escherichia coli by the recombinant vector pTrc99A-Sau-TBD and inducing protein expression.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.