title: TEV protease-mediated cleavage in Drosophila as a tool to analyze protein functions in living organisms.
abstract: Drosophila provides a powerful experimental system to analyze gene functions in a multi-cellular organism. Here we describe an in vivo method that interferes with the integrity of selected proteins through site-specific cleavage in Drosophila. The technique is based on the highly specific seven-amino-acid recognition site of the tobacco etch virus (TEV) protease. We established transgenic fly lines that direct TEV protease expression in various tissues without affecting fly viability. The insertion of the TEV protease recognition site in defined positions of target proteins mediates their sequence-specific cleavage after controlled TEV protease expression in the fly. Thereby, this technique is a powerful tool that allows the in vivo manipulation of selected proteins in a time- and tissue-specific manner.
authors.affiliation: Max-Planck-Institut für biophysikalische Chemie, Göttingen, Germany.
authors.forename: Ben
authors.initials: B
authors.lastname: Harder
authors.forename: Adrian
authors.initials: A
authors.lastname: Schomburg
authors.forename: Ralf
authors.initials: R
authors.lastname: Pflanz
authors.forename: Katharina
authors.initials: K
authors.lastname: Küstner
authors.forename: Nina
authors.initials: N
authors.lastname: Gerlach
authors.forename: Reinhard
authors.initials: R
authors.lastname: Schuh
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000378:metabolism*; D004330:Drosophila / Q000235:genetics* / Q000378:metabolism*; D029721:Drosophila Proteins / Q000502:physiology*; D010450:Endopeptidases / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*
chemical_list: D029721:Drosophila Proteins; D010450:Endopeptidases; C099488:TEV protease