Difference between revisions of "Part:BBa K2992026"
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | This parts entry represents an integration module for the expression of <i>botR</i> at the <i>pyrE</i> locus of the <i>C. sporogenes</i> genome. This module comprises the <i>botR</i> gene of <i>C. botulinum</i> [https://parts.igem.org/Part:BBa_K2992002 BBa_K2992002] coupled with its native RBS [https://parts.igem.org/Part:BBa_K2992011 BBa_K2992011] to encourage transcription from the promoter of the <i>pyrD</i> gene following integration onto the chromosome of <i>C. sporogenes</i>. A strong clostridial terminator was included to prevent polar transcription of <i> pyrE</i> and any downstream genes on the chromosome of <i>C. sporogenes</i> [https://parts.igem.org/Part:BBa_K2992013 BBa_K2992013]. In our project we use the transcriptional regulator of neurotoxin production from <i>C. botulinum</i>, BotR, to control the regulation of our volatile reporter operons (hyperlink to comp parts) and our fluorescent reporter FAST [https://parts.igem.org/Part:BBa_K2992000 BBa_K2992000] through interaction with its own promoter sequence P<i>botrR</i> [https://parts.igem.org/Part:BBa_K2992012 BBa_k299012] and P<i>ntnH</i> [https://parts.igem.org/Part:BBa_K2992001 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.<br><br> | + | This parts entry represents an integration module for the expression of <i>botR</i> at the <i>pyrE</i> locus of the <i>C. sporogenes</i> genome. This module comprises the <i>botR</i> gene of <i>C. botulinum</i> [https://parts.igem.org/Part:BBa_K2992002 BBa_K2992002] coupled with its native RBS [https://parts.igem.org/Part:BBa_K2992011 BBa_K2992011] to encourage transcription from the promoter of the <i>pyrD</i> gene following integration onto the chromosome of <i>C. sporogenes</i>. A strong clostridial terminator was included to prevent polar transcription of <i> pyrE</i> and any downstream genes on the chromosome of <i>C. sporogenes</i> [https://parts.igem.org/Part:BBa_K2992013 BBa_K2992013]. In our project we use the transcriptional regulator of neurotoxin production from <i>C. botulinum</i>, BotR, to control the regulation of our volatile reporter operons (hyperlink to comp parts) and our fluorescent reporter FAST [https://parts.igem.org/Part:BBa_K2992000 BBa_K2992000] through interaction with its own promoter sequence P<i>botrR</i> [https://parts.igem.org/Part:BBa_K2992012 BBa_k299012] and P<i>ntnH</i> [https://parts.igem.org/Part:BBa_K2992001 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.<br><br> |
===Characterisation=== | ===Characterisation=== | ||
Revision as of 10:39, 18 September 2019
botR integration module for C. sporogenes with native RBS for polar transcription
Usage and Biology
This parts entry represents an integration module for the expression of botR at the pyrE locus of the C. sporogenes genome. This module comprises the botR gene of C. botulinum BBa_K2992002 coupled with its native RBS BBa_K2992011 to encourage transcription from the promoter of the pyrD gene following integration onto the chromosome of C. sporogenes. A strong clostridial terminator was included to prevent polar transcription of pyrE and any downstream genes on the chromosome of C. sporogenes BBa_K2992013. In our project we use the transcriptional regulator of neurotoxin production from C. botulinum, BotR, to control the regulation of our volatile reporter operons (hyperlink to comp parts) and our fluorescent reporter FAST BBa_K2992000 through interaction with its own promoter sequence PbotrR BBa_k299012 and PntnH BBa_K2992001 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