Eukaryotic use of non-chimeric mutational vectors
US6010907A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | May 12, 1998 |
| Grant date | Jan 4, 2000 |
| Priority date | — |
| Expiry date | May 12, 2018 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N15/102
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The invention is based on the reaction of Duplex Mutational Vector in a cell-free system containing a cytoplasmic cell extract and a test plasmid. The reaction specifically converts a mutant kan.sup.r gene to recover the resistant phenotype in transformed MutS, RecA deficient bacteria. Using this system a type of Duplex Mutational Vector termed a Non-Chimeric Mutational Vector, having no RNA:DNA hybrid-duplex is shown to be an effective substrate for eukaryotic enzymes. The invention concerns the use of Non-Chimeric Mutational Vectors protected from 3' exonuclease attack in eukaryotic cells. Such protection can be conferred by replacement of a tetrathymidine linker by a nuclease resistant oligonucleotide, such as tetra-2'-O-methyl-uridine, to link the two strands of the recombinagenic oligonucleobase.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.