Part:BBa_K2992039
gusA reporter construct with PntnH 5-UTR+RBS
Usage and Biology
This parts entry represents a gusA reporter construct under the regulatory control of promoter for the non-toxic non-haemagglutinin ntnH gene of C. botulinum. The construct comprises PntnH BBa_K2992001 and its associated 5’-UTR and RBS driving the expression of the reporter gene gusA from E. coli BBa_K2300032 with a strong clostridial terminator TFad from C. pasteurianum BBa_K2284012. gusA encodes a B-glucorinidase from E. coli which can be used in a histochemical reaction to quantitatively measure promoter strength. This is accomplished through the addition of the fluorescent substrate 4 methylumbelliferyl glucuronide (4-MUG) which undergoes a colorimetric reaction with GusA, proportionate to its intracellular concentration. The nature of the Gus assay permits the measurement of promoter strength directly from cell debris without the requirement for extended cell lysis procedures. Doing so allows for the detection of reporter gene-products from anaerobic organisms which have recently been removed from the anaerobic cabinet or which have been frozen directly thereafter. In our project we use gusA as an established reporter gene for anaerobic organisms to validate the promoters chosen for our botR and acetone production constructs.
Characterisation
Data incoming.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
Heap Modular Joseph Recent Developments of the Synthetic Biology Toolkit for Clostridium
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