In vitro assay for inhibitors of the intron self-splicing reaction in Pneumocystis carinii
US5849484A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Jun 19, 1995 |
| Grant date | Dec 15, 1998 |
| Priority date | — |
| Expiry date | Jun 19, 2015 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC07K14/44
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention pertains to an in vitro method for assaying for an inhibitor of the catalytic Group I self-splicing intron reaction in the nuclear rRNA genes of Pheumocystis carinii which comprises the steps of (a) providing a DNA template containing the intron (I) from the 26S rRNA gene in Pneumocystis carinii and a portion of the 5' and 3' flanking exons (E1 and E2, respectively) between nucleotides 1963 and 2267 of 26S rRNA (660 nucleotides of amplified rRNA gene including the group I intron); (b) preparing an RNA precursor by transcription of the DNA template in the presence of labeled nucleoside triphosphates to produce a labeled RNA precursor (E1-I-E2); (c) purifying the RNA precursor; (d) incubating the RNA precursor and the inhibitor in the presence of guanosine triphosphate and magnesium ions; and (e) determining the degree of inhibition by the inhibitor on the intron splicing reaction in the RNA precursor by measuring the amount of labeled splicing intermediates and splicing products.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.