Method of increasing complementarity in a heteroduplex
US7235386B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jul 25, 2002 |
| Grant date | Jun 26, 2007 |
| Priority date | — |
| Expiry date | Oct 3, 2022 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N15/1027
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
We describe here an in vitro method of increasing complementarity in a heteroduplex polynucleotide sequence. The method uses annealing of opposite strands to form a polynucleotide duplex with mismatches. The heteroduplex polynucleotide is combined with an effective amount of enzymes having strand cleavage activity, 3′ to 5′ exonuclease activity, and polymerase activity, and allowing sufficient time for the percentage of complementarity to be increased within the heteroduplex. Not all heteroduplex polynucleotides will necessarily have all mismatches resolved to complementarity. The resulting polynucleotide is optionally ligated. Several variant polynucleotides result. At sites where either of the opposite strands has templated recoding in the other strand, the resulting percent complementarity of the heteroduplex polynucleotide sequence is increased. The parent polynucleotides need not be cleaved into fragments prior to annealing heterologous strands. Therefore, no reassembly is required.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.