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Updated: Aug 19, 2026

Generation of Fluorescent Protein Fusions in Candida Species
Published on: March 4, 2017
An improved GFP cloning cassette designed for prokaryotic transcriptional fusions
1Department of Plant and Microbial Biology, University of California, Berkeley 94720, USA.
Abstract:
A new gfp cloning cassette designed for prokaryotic transcriptional fusions has been constructed. This cassette consists of gfp (containing the S65T 'red-shift' [Heim et al. (1995) Nature 373, 663-664] and F64L 'protein solubility' [Cormack et al. (1996) Gene 173, 33-38] mutations) flanked by convenient restriction sites, a translational enhancer, and a consensus ribosome binding site with an optimized spacer region. gfp fusion strains containing this cassette demonstrate from 40- to 80-fold greater fluorescence intensity than wild-type gfp fusion strains. Additionally, this cassette confers sufficient fluorescence to recipient cells to be used in low copy-number plasmids, with promoters conferring low levels of transcription, and in bacterial taxa other than Escherichia, such as Pseudomonas.
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