Green fluorescent protein as a reporter of gene expression and protein localization

S. R. Kain, M. Adams, A. Kondepudi, T. T. Yang, W. W. Ward, P. Kitts

Research output: Contribution to journalArticlepeer-review

145 Scopus citations

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 (λ(max) = 509 nm) when excited with UV or blue light (λ(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(TM) 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 proteins to either the N or C terminus of GFP.

Original languageEnglish (US)
Pages (from-to)650-655
Number of pages6
JournalBioTechniques
Volume19
Issue number4
StatePublished - Jan 1 1995
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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