Bifunctional protein for the isolation of capped mRNA
US5219989A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Feb 24, 1992 |
| Grant date | Jun 15, 1993 |
| Priority date | — |
| Expiry date | Feb 24, 2012 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC07K2319/705
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A protein comprising at least a first functional site having the ability to bind the cap structure of mRNA and a second functional site having the ability to bind a solid support matrix in such a manner as to allow the first functional site to be immobilized and still remain functionally accessible to interact with the cap structure of mRNA. Also within the scope of the present invention is a method for generating a cDNA library mostly containing full-length cDNAs. The method comprises the incubation of a mixture comprising mRNA:cDNA hybrids with 1) a single strand RNA specific nuclease and 2) the above-mentioned protein. The resulting mixture is then passed through a column comprising a support matrix having the ability to bind the second functional site of the above-mentioned protein in order to selectively bind complete mRNA:cDNA hybrids. The mRNA:cDNA hybrids are then competitively eluted with a cap analog and full-length cDNA strands are separated and recovered. The present invention also includes a method for purifying capped mRNA using the above-mentioned protein. The process comprises the incubation of a mixture containing mRNA with the above-mentioned protein, passing the …
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.