Difference between revisions of "Part:BBa K2992025"
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===Usage and Biology=== | ===Usage and Biology=== | ||
− | This parts entry represents an integration module for the expression of <i>botR</i> from the <i>pyrE</i> locus of the <i>C. sporogenes</i> genome. This module comprises a strong clostridial terminator sequence T<i>fad</i> to prevent polar transcription from <i>pyrD</i> [https://parts.igem.org/Part:BBa_K2992013] and the <i>botR gene of<i>C. botulinum</i> [https://parts.igem.org/Part:BBa_K2992002], under the regulatory control of its native promoter P<i>botR</i> [https://parts.igem.org/wiki/Part:BBa_K2992012] and its associated 5’-UTR which contains the RBS [https://parts.igem.org/Part:BBa_K2992014]. An additional strong clostridial terminator was included to prevent polar effects for the transcription of<i> pyrE</i> and any downstream genes on the chromosome of <i>C. sporogenes. In our project we use the transcriptional regulator BotR to control the regulation of our volatile reporter operons | + | This parts entry represents an integration module for the expression of <i>botR</i> from the <i>pyrE</i> locus of the <i>C. sporogenes</i> genome. This module comprises a strong clostridial terminator sequence T<i>fad</i> to prevent polar transcription from <i>pyrD</i> [https://parts.igem.org/Part:BBa_K2992013] and the <i>botR gene of<i>C. botulinum</i> [https://parts.igem.org/Part:BBa_K2992002], under the regulatory control of its native promoter P<i>botR</i> [https://parts.igem.org/wiki/Part:BBa_K2992012] and its associated 5’-UTR which contains the RBS [https://parts.igem.org/Part:BBa_K2992014]. An additional strong clostridial terminator was included to prevent polar effects for the transcription of<i> pyrE</i> and any downstream genes on the chromosome of <i>C. sporogenes. In our project we use the transcriptional regulator BotR to control the regulation of our volatile reporter operons (add comp part hyperlinks) and our fluorescent reporter FAST [] through interaction with its own promoter sequence P<i>botrR</i> [https://parts.igem.org/Part:BBa_K2992012] and P<i>ntnH</i> [https://parts.igem.org/Part:BBa_K2992001] whose genes are cognate members of the BotR regulon. Doing so allows us to use our surrogate host strain <i>C. sporogenes</i> as a model system for predicting botulinum neurotoxin production following food manufacture, through the detection of our chosen reporters. |
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Revision as of 08:16, 18 September 2019
botR integration module for C. sporogenes with native promoter, 5-UTR+RBS
Usage and Biology
This parts entry represents an integration module for the expression of botR from the pyrE locus of the C. sporogenes genome. This module comprises a strong clostridial terminator sequence Tfad to prevent polar transcription from pyrD [1] and the botR gene of<i>C. botulinum [2], under the regulatory control of its native promoter PbotR [3] and its associated 5’-UTR which contains the RBS [4]. An additional strong clostridial terminator was included to prevent polar effects for the transcription of pyrE and any downstream genes on the chromosome of C. sporogenes. In our project we use the transcriptional regulator BotR to control the regulation of our volatile reporter operons (add comp part hyperlinks) and our fluorescent reporter FAST [] through interaction with its own promoter sequence P<i>botrR [5] and PntnH [6] whose genes are cognate members of the BotR regulon. Doing so allows us to use our surrogate host strain C. sporogenes as a model system for predicting botulinum neurotoxin production following food manufacture, through the detection of our chosen reporters.
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
Cañadas et al., 2019 RiboCas – update Dupuy and Matamouros, 2006 update Minton et al., 2016 Road map – update Raffestin et al 2009 - update Dupuy and Matamouros, 2006 update