Method for generating recombinant DNA library using unidirectional single-stranded DNA fragments
US6955879B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jun 16, 2001 |
| Grant date | Oct 18, 2005 |
| Priority date | — |
| Expiry date | Apr 16, 2023 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N15/1027
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention relates to a method for producing a recombinant polynucleotides comprising the steps of generating a pool of unidirectional single-stranded polynucleotide fragments from two or more homologous double-stranded polynucleotides, conducting a polymerization process comprising multi-cyclic extension reactions using the unidirectional single-stranded polynucleotide fragments as templates and specific oligonucleotides as primers to obtain recombinant polynucleotides, and conducting a polymerase chain reaction using at least one primer to amplify the recombinant polynucleotides; and a method for constructing a recombinant DNA library comprising the steps of inserting the recombinant polynucleotide prepared by the above method into a vector and transforming an expression cell with said vector containing the recombinant polynucleotide to obtain a plurality of mutant clones. The method of the present invention can be used for in vitro recombination of homologous polynucleotides and the directed molecular evolution.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.