Nucleic acids encoding oxidation-reduction sensitive green fluorescent protein variants

US7314736B2 · kind B2 · utility

Assignee

• The State of Oregon Acting by and through the State Board of Higher Education on Behalf of the University of Oregon · US

Inventors

• S. James Remington · Eugene, US
• George Hanson · Eugene, US

Key dates

• Filing date: Oct 20, 2005
• Grant date: Jan 1, 2008
• Expiry date: Feb 7, 2026

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C07K14/43595
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The disclosure provides proteins that can be used to determine the redox status of an environment (such as the environment within a cell or subcellular compartment). These proteins are green fluorescent protein (GFP) variants (also referred to as redox sensitive GFP (rosGFP) mutants), which have been engineered to have two cysteine amino acids near the chromophore and within disulfide bonding distance of each other. Also provided are nucleic acid molecules that encode rosGFPs, vectors containing such encoding molecules, and cells transformed therewith. The disclosure further provides methods of using the rosGFPs (and encoding molecules) to analyze the redox status of an environment, such as a cell, or a subcellular compartment within a cell. In certain embodiments, both redox status and pH are analyzed concurrently.