Difference between revisions of "Part:BBa K2992047"
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | This parts entry represents a FAST reporter construct under the regulatory control of P<i>ha33</i> for the hemagglutinin component of the Botulinum neurotoxin complex in <i>C. botulinum</i>. The construct comprises the promoter P<i>ha33</i> [https://parts.igem.org/Part:BBa_K2992040 BBa_K2992040] coupled with its associated 5’-UTR containing the RBS [https://parts.igem.org/Part:BBa_K272992041 BBa_K2992041], driving the expression of the Fluorescence-Activating and Absorption-Shifting Tag protein (FAST) reporter gene [http//parts.igem.org/Part:BBa_K2992000 BBa_K2992000]. Transcirptional terminator occurs through the activity of the strong clostridial terminator T<i>Fad</i> [https://parts.igem.org/Part:BBa_K2284012 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 <i>Clostridium</i>. 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. <br><br> | + | This parts entry represents a FAST reporter construct under the regulatory control of P<i>ha33</i> for the hemagglutinin component of the Botulinum neurotoxin complex in <i>C. botulinum</i>. The construct comprises the promoter P<i>ha33</i> [https://parts.igem.org/Part:BBa_K2992040 BBa_K2992040] coupled with its associated 5’-UTR containing the RBS [https://parts.igem.org/Part:BBa_K272992041 BBa_K2992041], driving the expression of the Fluorescence-Activating and Absorption-Shifting Tag protein (FAST) reporter gene [http//parts.igem.org/Part:BBa_K2992000 BBa_K2992000]. Transcirptional terminator occurs through the activity of the strong clostridial terminator T<i>Fad</i> [https://parts.igem.org/Part:BBa_K2284012 BBa_K2284012]. FAST is one of the few fluorescent reporters available for effective use in anaerobic organisms. FAST is derived from <i>Halorhodospira halophila</i> and has been codon optimised for fluorescence studies in the genus <i>Clostridium</i>. 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. <br><br> |
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Revision as of 13:10, 1 October 2019
FAST reporter construct with Pha33 5-UTR+RBS
Usage and Biology
This parts entry represents a FAST reporter construct under the regulatory control of Pha33 for the hemagglutinin component of the Botulinum neurotoxin complex in C. botulinum. The construct comprises the promoter Pha33 BBa_K2992040 coupled with its associated 5’-UTR containing the RBS BBa_K2992041, driving the expression of the Fluorescence-Activating and Absorption-Shifting Tag protein (FAST) reporter gene [http//parts.igem.org/Part:BBa_K2992000 BBa_K2992000]. Transcirptional 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 276
References
Heap modular Street et al 2019. Update