Prokaryotic two-hybrid system

US6051381A · kind A · utility

Inventor

• Michael Kornacker · Seattle, US

Key dates

• Filing date: Dec 10, 1997
• Grant date: Apr 18, 2000
• Expiry date: Dec 10, 2017

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C07K2319/00
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A two-hybrid system that can detect homo- and heterodimeric protein interactions in E. coli and other cells. This system is useful for the same applications as a yeast two-hybrid system; i.e. interaction cloning, mapping protein interaction domains, analyzing protein interactions, detecting protein interactions and detecting modulators thereof. The invention concerns a prokaryotic host cell comprising: PA0 (a) a fusion protein having (i) a first DNA-binding domain and (ii) a first interacting domain; PA0 (b) a fusion protein having (i) a second DNA-binding domain and (ii) a second interacting domain capable of binding to the first interacting domain; and PA0 (c) a nucleic acid molecule having a reporter gene operatively linked to (i) a promoter, (ii) a first operator site capable of binding to the first DNA-binding domain, located upstream of the promoter, and (iii) a second operator site capable of binding the second DNA-binding domain, located downstream of the promoter of the reporter gene; wherein binding of the first interacting domain to the second interacting domain is signaled by altered expression of the reporter gene.