System to detect protein-protein interactions

US5283173A · kind A · utility

Assignee

• The Research Foundation of the State University of New York · US

Inventors

• Stanley Fields · East Setauket, US
• Ok-Kyu Song · Stony Brook, US

Key dates

• Filing date: Jan 24, 1990
• Grant date: Feb 1, 1994
• Expiry date: Jan 24, 2010

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N15/1055
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Methods are provided for detecting the interaction between a first test protein and a second test protein, in vivo, using reconstitution of the activity of a transcriptional activator. This reconstitution makes use of chimeric genes which express hybrid proteins. Two types of hybrid proteins are prepared. The first hybrid contains the DNA-binding domain of a transcriptional activator fused to the first test protein. The second hybrid protein contains a transcriptional activation domain fused to the second test protein. If the two test proteins are able to interact, they bring into close proximity the two domains of the transcriptional activator. This proximity is sufficient to cause transcription, which can be detected by the activity of a marker gene which contains a binding site for the DNA-binding domain.