Green Fluorescent Protein FP506
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Fluorescent proteins have become widely used in a variety of experimental paradigms since the demonstration of the utility of the first one, Green Fluorescent Protein (GFP) about 20 years ago (1). In 1999 Matz et al. characterized several fluorescent proteins from various coral species which were cloned and sequenced using degenerate primers based on the GFP sequence (2). One of these was Fp506, isolated from a
coral. This protein proved to have spectral properties similar to GFP although the protein sequence was only distantly related.
His-tagged-Fp506 was run out on an SDS-PAGE gel at 6µg and 3µg in the indicated lanes in the middle of the SDA-PAGE gel. BSA was also run at 6µg and 3µg per lane as indicated. The first lane contains SDS-PAGE standards of indicated molecular size. The pET29a vector adds a C-terminal His-tag which was use to purify the protein and this, along with some other vector derived sequence, adds about 5 kDa to the molecule, resulting in a ~32kDa protein. The left two lanes show similar preparations of another fluorescent protein, the photoconvertible green-red protein EosFP which is a little smaller in molecular size than Fp506
Our fp506 construct is identical to that in the Uniprot database entry
, as described in reference 1. The vector adds a C-terminal His-tag which was use to purify the protein and this, along with some other vector derived sequence, adds about 5 kDa to the molecule.
1. Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC. Green fluorescent protein as a marker for gene expression.
Science 263:802-5 (1994).
2. Matz MV, Fradkov AF, Labas YA, Savitsky AP, Zaraisky AG, Markelov ML and Lukyanov SA. Fluorescent proteins from nonbiolu.minescent Anthozoa species
Nat. Biotechnol. 17: 969-973 (1999).