Methods and materials for the rapid and high volume production of a gene knock-out library in an organism
US6703200B1 · kind B1 · utility
Assignee
Inventors
Key dates
| Filing date | Sep 11, 2000 |
| Grant date | Mar 9, 2004 |
| Priority date | — |
| Expiry date | Sep 11, 2020 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY02T10/70
- WIPO fieldElectrical machinery, apparatus, energy
- WIPO sectorElectrical engineering
Abstract
The present invention relates to a method for facilitating site directed homologous recombination in an organism to produce mutants comprising:1) providing a large insert vector library comprising one or more large insert vectors, each of said large insert vectors comprising a piece of DNA, said DNA piece comprising multiple genes from a target organism and a first selectable marker functional for selection in bacteria;2) providing a second vector comprising a transposable element, said transposable element comprising a nucleotide sequence coding for a second selectable marker flanked on each side by an inverted repeat sequence, wherein said selectable marker is bifunctional for selection in bacteria and the target organism and wherein said inverted repeat sequences are functional as a binding site for a transposase;3) incubating said library with said second vector in the presence of a transposase specific for the inverted repeat sequences on the plasmid vector, such that the transposable element is transferred randomly into an individual large insert of the large insert library to produce disrupted large insert vectors;4) optionally, amplifying the disrupted large insert vectors …
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.