Difference between revisions of "Part:BBa K2992045"

Latest revision as of 13:45, 21 October 2019

FAST reporter construct with Pthl 5-UTR+RBS.

Usage and Biology

This parts entry represents a FAST reporter construct under the regulatory control of Ptthl for the thiolase gene of C. acetobutylicum. The construct comprises the strong clostridial promoter Pthl K2715010BBa_ K2715010 coupled with its associated 5’-UTR K2715019 BBa_ K2715019 and RBS BBa_K2992009, driving the expression of the fluorescent reporter gene FAST BBa_K2992000. Transcriptional terminator occurs through the activity of the strong clostridial terminator TFad BBa_K2284012. FAST is one of the few fluorescent reporters available for effective use in anaerobic organisms. FAST is derived from Halorhodospira halophila and has been codon optimised for fluorescence studies in the genus Clostridium. In our project we couple FAST with the natural promoters of the BotR regulon thus linking reporter fluorescence with botulinum neurotoxin production. In doing so, we hoped to generate our surrogate host strain as a model for predicting neurotoxin production in foodstuffs following food manufacturing processes.

Characterisation

Data incoming.

Sequence and Features

References

Streett, H., Kalis, K. and Papoutsakis, E. (2019). A Strongly Fluorescing Anaerobic Reporter and Protein-Tagging System for Clostridium Organisms Based on the Fluorescence-Activating and Absorption-Shifting Tag Protein (FAST). Applied and Environmental Microbiology, 85(14).

Heap, J., Pennington, O., Cartman, S. and Minton, N. (2009). A modular system for Clostridium shuttle plasmids. Journal of Microbiological Methods, 78(1), pp.79-85.