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Green Fluorescent Protein as a Model for Protein Crystal Growth StudiesGreen fluorescent protein (GFP) from jellyfish Aequorea Victoria has become a popular marker for e.g. mutagenesis work. Its fluorescent property, which originates from a chromophore located in the center of the molecule, makes it widely applicable as a research too]. GFP clones have been produced with a variety of spectral properties, such as blue and yellow emitting species. The protein is a single chain of molecular weight 27 kDa and its structure has been determined at 1.9 Angstrom resolution. The combination of GFP's fluorescent property, the knowledge of its several crystallization conditions, and its increasing use in biophysical and biochemical studies, all led us to consider it as a model material for macromolecular crystal growth studies. Initial preparations of GFP were from E.coli with yields of approximately 5 mg/L of culture media. Current yields are now in the 50 - 120 mg/L range, and we hope to further increase this by expression of the GFP gene in the Pichia system. The results of these efforts and of preliminary crystal growth studies will be presented.
Document ID
19980236877
Document Type
Reprint (Version printed in journal)
Authors
Agena, Sabine (London Univ. United Kingdom)
Smith, Lori (Alabama Univ. Huntsville, AL United States)
Karr, Laurel (NASA Marshall Space Flight Center Huntsville, AL United States)
Pusey, Marc (NASA Marshall Space Flight Center Huntsville, AL United States)
Date Acquired
August 18, 2013
Publication Date
January 1, 1998
Subject Category
Solid-State Physics
Meeting Information
Crystallization of Biological Maromolecules(Granada)
Distribution Limits
Public
Copyright
Work of the US Gov. Public Use Permitted.