Expression library screen by prenylation of expressed proteins
US5932432A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Jan 21, 1997 |
| Grant date | Aug 3, 1999 |
| Priority date | — |
| Expiry date | Jan 21, 2017 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/48
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Described is a novel in vitro method for obtaining and identifying proteins which, in their natural in vivo setting, are covalently modified after translation. To identify novel isoprenylated proteins for subsequent biochemical study, colony blots of a Glycine max cDNA expression library were .sup.3 H! farnesyl labeled in vitro. Proteins identified by this screen contained several different carboxy-termini that conform to consensus farnesylation motifs. These proteins included known farnesylated proteins (DnaJ homologs) and several novel proteins, two of which contained 6 or more tandem repeats of a hexapeptide having the consensus sequence E or G! G or P!EKP or K!K. Expression library screening by direct labeling can thus be adapted to recover and identify isoprenylated proteins as well as proteins with other post-translational modifications. This identification and recovery further enables the recovery of transformants containing DNA encoding the proteins, as well as the raising of antibodies to the proteins.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.