Mutant Neq HS DNA polymerase derived from Nanoarchaeum equitans and its application to hot-start PCR
US9416352B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Nov 28, 2014 |
| Grant date | Aug 16, 2016 |
| Priority date | — |
| Expiry date | Nov 28, 2034 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC07K2319/21
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A DNA polymerase (Neq DNA polymerase) derived from Nanoarchaeum equitans is split into Neq L and Neq S fragments, each of which contains inteins. A Neq hot-start (HS) DNA polymerase in which the inteins of the Neq L and Neq S fragments are linked with each other is provided in the form of a precursor of Neq DNA polymerase. A purification method can be significantly improved by inserting a His-tag sequence composed of six histidine residues between the inteins of the Neq L and Neq S fragments at a gene level. As a result of effort to enhance PCR efficiency of the Neq HS DNA polymerase, a gene coding for the Neq HS DNA polymerase is mutated at specific positions to screen mutant Neq HS polymerases (M1, M2, and M3) having a highly improved PCR amplification rate and amplification level.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.