Rapid selection against truncation mutants in yeast reverse two-hybrid screens

Rapid selection against truncation mutants in yeast reverse two-hybrid screens

The yeast reverse two-hybrid system is a powerful technique for isolating mutations in a protein that abolish its interaction with a known partner. Selection is based on abrogation of growth suppression imposed when wild-type interactions confer 5-fluoroorotic acid (5-FOA) sensitivity to yeast cells...

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Veröffentlicht in:BioTechniques. - 1993. - 30(2001), 5 vom: 01. Mai, Seite 984-8
1. Verfasser: Puthalakath, H (VerfasserIn)
Weitere Verfasser: Strasser, A, Huang, D C
Format: Aufsatz
Sprache:English
Veröffentlicht: 2001
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Technical Report Journal Article Apoptosis Regulatory Proteins Bcl-2-Like Protein 11 Carrier Proteins Drosophila Proteins Luminescent Proteins Membrane Proteins mehr... Proto-Oncogene Proteins Recombinant Fusion Proteins Green Fluorescent Proteins 147336-22-9 Orotic Acid 61H4T033E5 5-fluoroorotic acid 7IA9OUC93E beta-Galactosidase EC 3.2.1.23 Dyneins EC 3.6.4.2 Fluorouracil U3P01618RT
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500 |a Date Completed 04.12.2001 
500 |a Date Revised 28.09.2018 
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500 |a ErratumIn: Biotechniques 2001 Sep;31(3):488 
500 |a Citation Status MEDLINE 
520 |a The yeast reverse two-hybrid system is a powerful technique for isolating mutations in a protein that abolish its interaction with a known partner. Selection is based on abrogation of growth suppression imposed when wild-type interactions confer 5-fluoroorotic acid (5-FOA) sensitivity to yeast cells. A laborious component of this system is to eliminate those mutations that cause protein truncation. By fusing the green fluorescent protein (GFP) to the C-terminus of a protein of interest, dynein light chain (LC8), we were able to rapidly isolate mutations that did not result in protein truncation 
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