title: Green fluorescent protein as a reporter of gene expression and protein localization.
abstract: The green fluorescent protein (GFP) from the jellyfish Aequorea victoria is rapidly becoming an important reporter molecule for monitoring gene expression and protein localization in vivo, in situ and in real time. GFP emits bright green light (lambda max = 509 nm) when excited with UV or blue light (lambda max = 395 nm, minor peak at 470 nm). The fluorescence excitation and emission spectra of GFP are similar to those of fluorescein, and the conditions used to visualize this fluorophore are also suitable for GFP. Unlike other bioluminescent reporters, the chromophore in GFP is intrinsic to the primary structure of the protein, and GFP fluorescence does not require a substrate or cofactor. GFP fluorescence is stable, species-independent and can be monitored non-invasively in living cells and, in the case of transparent organisms, whole animals. Here we demonstrate GFP fluorescence in bacterial and mammalian cells and introduce our Living Colors line of GFP reporter vectors, GFP protein and anti-GFP antiserum. The reporter vectors for GFP include a promoterless GFP vector for monitoring the expression of cloned promoters/enhancers in mammalian cells and a series of six vectors for creating fusion protein to either the N or C terminus of GFP.
authors.affiliation: CLONTECH Laboratories, Inc., Palo Alto, CA, USA.
authors.forename: S R
authors.initials: SR
authors.lastname: Kain
authors.forename: M
authors.initials: M
authors.lastname: Adams
authors.forename: A
authors.initials: A
authors.lastname: Kondepudi
authors.forename: T T
authors.initials: TT
authors.lastname: Yang
authors.forename: W W
authors.initials: WW
authors.lastname: Ward
authors.forename: P
authors.initials: P
authors.lastname: Kitts
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D015870:Gene Expression; D005819:Genetic Markers; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins; D015183:Restriction Mapping; D012486:Salmonella typhimurium / Q000235:genetics; D013050:Spectrometry, Fluorescence; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D005819:Genetic Markers; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins