Green Fluorescent Protein, Recombinant
Aequoria coerulescens GFP, AcGFP
This protein originates from an Aequoria coerulescens, a close relative of A. victoria, and the protein was engineered to improve spectral properties and prevent oligomerization. This form of GFP, referred to as AcGFP, is 94% identical to the eGFP developed by Tsien and coworkers and is the form of GFP inserted in the Clontech/Takara pAcGFP and related expression vectors.
The green fluorescent protein (GFP) is a 27kD protein isolated originally from the jellyfish Aequoria victoria. It has an endogenous fluorochrome activity with excitation maximum at 395nm and emission maximum at 509nm, which is similar to that of fluorescein (1,2). The GFP gene was cloned and sequenced and the origin of the fluorochrome by autocatalytic activity of certain amino acids was discovered (3,4). Much interest in GFP was generated when it was shown that fluorescence develops rapidly when the protein is expressed and requires only molecular oxygen and no other cofactors. As a result GFP can be expressed in fluorescent form in essentially any prokaryotic or eukaryotic cell (5). GFP has been engineered to produce a vast number of variously colored mutants including blue, cyan and yellow protein derivatives, BFP, CFP and YFP (6-9). GFP and other fluorescent proteins derived from jellyfish, coral and other Cnidaria are widely used as tracers in transfection and transgenic experiments to monitor gene expression and protein localization in vivo and in in vitro. The crystal structure of GFP was determined (7) which allowed amino acid modifications to improve spectral properties and prevent multimerization (8,9). The 2008 Nobel prize in chemistry was awarded “for the discovery and development of the green fluorescent protein, GFP”.
Recombinant protein corresponding to Aequoria coerulescens GFP, fused to His-Tag at C-terminal, expressed in E. coli. Swiss/Uniprot: Q6YGZ0
Amino Acid Sequence
MKETAAAKFE RQHMDSPDLG TLVPRGSMAD IGSEFMVSKG AELFTGIVPI LIELNGDVNG HKFSVSGEGE GDATYGKLTL KFICTTGKLP VPWPTLVTTL SYGVQCFSRY PDHMKQHDFF KSAMPEGYIQ ERTIFFEDDG NYKSRAEVKF EGDTLVNRIE LTGTDFKEDG NILGNKMEYN YNAHNVYIMT DKAKNGIKVN FKIRHNIEDG SVQLADHYQQ NTPIGDGPVL LPDNHYLSTQ SALSKDPNEK RDHMIYFGFV TAAAITHGMD ELYKVDKLAAALE HHHHHH
Applications
Suitable for use as a fluorescent protein standard and as an immunogen for antibody production. Other applications not tested.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing.. Store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Source
Recombinant, E. coli
Form
Supplied as a liquid in 6M urea, 10mM phosphate, pH 7.5
Important Note
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
