DNA recombination
US10876125B2 · kind B2 · utility
Assignees
Inventors
Key dates
| Filing date | Aug 5, 2016 |
| Grant date | Dec 29, 2020 |
| Priority date | — |
| Expiry date | Aug 12, 2036 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N2760/14134
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
There is provided a non-PCR based method for construction of a DNA recombination fragment with necessary flanking regions homologous to a desired site of genetic manipulation within a target DNA required for recombination-based manipulation of said target DNA, comprising the steps of: A) identifying a desired site of insertion for a genetic element in said target DNA; B) locating an endogenous, native, half-site of a selected restriction endonuclease present upstream of the site within the DNA to be targeted for genetic manipulation and thereby defining the 5′ extent of an upstream homology region; C) locating an endogenous, native, half-site of the same selected restriction endonuclease present downstream of the site within the DNA to be targeted for genetic manipulation and thereby defining the 3′ extent of a downstream homology region; D) synthesising a flanking region cassette resulting in juxtapositioning of the upstream and downstream half-sites thereby causing formation of complete restriction site for the selected restriction endonuclease flanked by said upstream and downstream homology regions; and E) inserting the flanking region cassette into a plasmid vector which inclu…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.