Brenda J Biermann

US Patent:

59324325, Aug 3, 1999

Filed:

Jan 21, 1997

Appl. No.:

8/785795

Inventors:

Dring N. Crowell - Indianapolis IN
Brenda Biermann - Zionsville IN
Stephen Randall - Speedway IN

Assignee:

Indiana University Foundation - Bloomington IN

International Classification:

C12Q 148
C12Q 168
G01N 3358

US Classification:

435 15

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!EK�P 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.