Recombination of polynucleotide sequences using random or defined primers
US6153410A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Aug 4, 1997 |
| Grant date | Nov 28, 2000 |
| Priority date | — |
| Expiry date | Aug 4, 2017 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/683
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method for in vitro mutagenesis and recombination of polynucleotide sequences based on polymerase-catalyzed extension of primer oligonucleotides is disclosed. The method involves priming template polynucleotide(s) with random-sequences or defined-sequence primers to generate a pool of short DNA fragments with a low level of point mutations. The DNA fragments are subjected to denaturization followed by annealing and further enzyme-catalyzed DNA polymerization. This procedure is repeated a sufficient number of times to produce full-length genes which comprise mutants of the original template polynucleotides. These genes can be further amplified by the polymerase chain reaction and cloned into a vector for expression of the encoded proteins.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.