PLEF, A NOVEL VECTOR FOR EXPRESSION OF GLUTATHIONE-S-TRANSFERASE FUSION PROTEINS IN MAMMALIAN-CELLS

Authors
RUDERT F VISSER E GRADL G GRANDISON P SHEMSHEDINI L WANG Y GRIERSON A WATSON J
Citation
F. Rudert et al., PLEF, A NOVEL VECTOR FOR EXPRESSION OF GLUTATHIONE-S-TRANSFERASE FUSION PROTEINS IN MAMMALIAN-CELLS, Gene, 169(2), 1996, pp. 281-282
Citations number
7
Categorie Soggetti
Genetics & Heredity
Journal title
Gene → ACNP
ISSN journal
03781119
Volume
169
Issue
2
Year of publication
1996
Pages
281 - 282
Database
ISI
SICI code
0378-1119(1996)169:2<281:PANVFE>2.0.ZU;2-6
Abstract
An expression vector, pLEF, has been used to produce the intracellular domain (IC) of the human CD95 (Fas/APO-1) apoptosis receptor as a glu tathione S-transferase (GST) fusion protein in murine L929 fibroblasts . GST::CD95IC was affinity-purified in a single step using glutathione -Sepharose. Purification of GST::CD95IC from P-32-labelled L999 cells and cleavage with thrombin revealed that CD95IC was phosphorylated in vivo when produced as a GST fusion protein. Therefore, pLEF may facili tate the mapping of in vivo-modified sites of eukaryotic proteins.