Green fluorescent protein TurboGFP

- Extra fast protein maturation
- Bright green fluorescence
- Proven suitability to generate stably transfected cell lines
- Efficient maturation at a wide range of temperatures

Available variants and fusions

TurboGFP codon usage is optimized for high expression in mammalian cells [Haas et al., 1996], but it can be successfully expressed in other heterological systems.

TurboGFP-mito fusion: A mitochondrial targeting sequence (MTS) is linked to the TurboGFP N-terminus. MTS was derived from the subunit VIII of human cytochrome C oxidase [Rizzuto et al., 1989; Rizzuto et al., 1995]. When expressed in mammalian cells, this variant provides green fluorescent labeling of mitochondria.

Destabilized TurboGFP variant (TurboGFP-dest1): TurboGFP-dest1 is produced by fusing the initial protein with PEST amino acid sequence encoded by region 422-461 of mouse ornithine decarboxylase gene [Li et al., 1998]. This sequence targets the protein to degradation and enables a rapid protein turnover. TurboGFP-dest1 retains spectral properties of the initial protein, but has shorter half-life (approximately 2 hrs) as measured by the analysis of fluorescence intensity of cells treated with a protein synthesis inhibitor, cycloheximide. Because of rapid turnover, TurboGFP-dest1 can be used to measure changes in gene expression.

Fluorescence intensities of cells expressing TurboGFP-dest1 rapidly decrease in response to cycloheximide (CHX). Mammalian cells expressing TurboGFP-dest1 under the control of CMV promoter were treated with CHX. After 1.5 hours CHX treatment, fluorescence intensity of cells was greatly reduced. Control cells - no CHX treatment.