Screening and use of agents which block or activate intein splicing utilizing natural or homologous exteins
US7157224B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Dec 18, 2002 |
| Grant date | Jan 2, 2007 |
| Priority date | — |
| Expiry date | Feb 26, 2024 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2333/35
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
In accordance with the present invention, there are provided selection systems and methods for screening for agents that control splicing of inteins in their native host protein (extein) or in homologous exteins. Specifically, there are provided positive genetic selection systems for the screening of agents which inhibit or activate protein splicing which comprise: a host cell containing a chromosomal gene encoding either a drug-resistant form of a target enzyme or a wild-type target enzyme, and a plasmid-borne gene encoding either a drug-sensitive form of the target enzyme, which is dominantly cytotoxic upon interaction with the drug, or a dominantly cytotoxic form of the target enzyme. In these systems the plasmid-borne gene contains an intein, and the inhibition or activation of splicing of the dominant cytotoxic form of the target enzyme by a given reagent results in the survival or death of the host cell. More specifically, positive genetic selection systems which utilize the M. xenopi GyrA intein or M. tuberculosis DnaB helicase intein are provided. Similar reporter systems utilizing native or homologous exteins and systems utilizing controllable inteins are provided, as are …
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.