Difference between revisions of "Part:BBa K2992030"
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===Usage and Biology===
 
===Usage and Biology===
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This parts entry represents an acetone-production pathway for plasmid-borne expression in <i>C. sporogenes</i> for predicting botulinum neurotoxin production. The entry comprises the thiolase gene <i>thl</i> [https://parts.igem.org/Part:BBa_K2992008 BBa_K2992008] and acetoacetate decarboxylase <i>adc</i> gene of <i>C. acetobutylicum</i> [https://parts.igem.org/Part:BBa_M36585 BBa_M36585] coupled with the two units of the <i>ctfAB</i> complex from <i>C. botulinum</i> [https://parts.igem.org/Part:BBa_K2992003 BBa_K2992003] and [https://parts.igem.org/Part:BBa_K2992005 BBa_K2992005] separated by their native intergenic region containing a partial RBS sequence for <i>ctfB</i> [https://parts.igem.org/Part:BBa_K2992007 BBa_K2992007]. This operon is regulated by the promoter [https://parts.igem.org/Part:BBa_K2992016 BBa_K2992016] and associated 5’UTR+RBS [https://parts.igem.org/Part:BBa_K2992017 BBa_K2992017] from the ferredoxin <i>fdx</i> gene of <i>C. sporogenes</i>.Transcriptional termination for this synthetic acetone-production operon occurs through the activity of T<i>fdx</i> from <i>C. pasteurianum</i> [https://parts.igem.org/Part:BBa_ K2284012 BBa_K2284012]. In our project we used genome-scale modelling to predict the necessary genes required to produce acetone in our chosen surrogate strain <i>C. sporogenes</i>. We sought to link acetone production with <i>C. botulinum</i> neurotoxin production by the integration of the neurotoxin transcriptional regulator <i>botR</i> onto the chromosome of <i>C. sporogenes</i> and by using promoter regions from the regulon of <i>botR</i> to control the acetone-production operons.  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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This parts entry represents an acetone-production pathway for plasmid-borne expression in <i>C. sporogenes</i> for predicting Botulinum neurotoxin production. The entry comprises the thiolase gene <i>thl</i> [https://parts.igem.org/Part:BBa_K2992008 BBa_K2992008] and acetoacetate decarboxylase <i>adc</i> gene of <i>C. acetobutylicum</i> [https://parts.igem.org/Part:BBa_M36585 BBa_M36585] coupled with the two units of the <i>ctfAB</i> complex from <i>C. botulinum</i> [https://parts.igem.org/Part:BBa_K2992003 BBa_K2992003] and [https://parts.igem.org/Part:BBa_K2992005 BBa_K2992005] separated by their native intergenic region containing a partial RBS sequence for <i>ctfB</i> [https://parts.igem.org/Part:BBa_K2992007 BBa_K2992007]. This operon is regulated by the promoter [https://parts.igem.org/Part:BBa_K2992016 BBa_K2992016] and associated 5’UTR+RBS [https://parts.igem.org/Part:BBa_K2992017 BBa_K2992017] from the ferredoxin <i>fdx</i> gene of <i>C. sporogenes</i>.Transcriptional termination for this synthetic acetone-production operon occurs through the activity of T<i>fdx</i> from <i>C. pasteurianum</i> [https://parts.igem.org/Part:BBa_ K2284012 BBa_K2284012]. In our project we used genome-scale modelling to predict the necessary genes required to produce acetone in our chosen surrogate strain <i>C. sporogenes</i>. We sought to link acetone production with <i>C. botulinum</i> neurotoxin production by the integration of the neurotoxin transcriptional regulator <i>botR</i> onto the chromosome of <i>C. sporogenes</i> and by using promoter regions from the regulon of <i>botR</i> to control the acetone-production operons.  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 09:06, 25 September 2019


Acetone pathway: csp_Pfdx-5-UTR+RBS-ca_thl-cb_ctfAB-cp_TFdx

Botulinum toxin-predicting acetone production pathway with C. sporogenes Pfdx driving expression of ctfAB and theC. acetobutylicum genes thl and adc.

Usage and Biology

This parts entry represents an acetone-production pathway for plasmid-borne expression in C. sporogenes for predicting Botulinum neurotoxin production. The entry comprises the thiolase gene thl BBa_K2992008 and acetoacetate decarboxylase adc gene of C. acetobutylicum BBa_M36585 coupled with the two units of the ctfAB complex from C. botulinum BBa_K2992003 and BBa_K2992005 separated by their native intergenic region containing a partial RBS sequence for ctfB BBa_K2992007. This operon is regulated by the promoter BBa_K2992016 and associated 5’UTR+RBS BBa_K2992017 from the ferredoxin fdx gene of C. sporogenes.Transcriptional termination for this synthetic acetone-production operon occurs through the activity of Tfdx from C. pasteurianum K2284012 BBa_K2284012. In our project we used genome-scale modelling to predict the necessary genes required to produce acetone in our chosen surrogate strain C. sporogenes. We sought to link acetone production with C. botulinum neurotoxin production by the integration of the neurotoxin transcriptional regulator botR onto the chromosome of C. sporogenes and by using promoter regions from the regulon of botR to control the acetone-production operons. 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


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 569
    Illegal PstI site found at 3464
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 3464
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 569
    Illegal PstI site found at 3464
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 569
    Illegal PstI site found at 3464
  • 1000
    COMPATIBLE WITH RFC[1000]

References

Heap Modular Cornillo et al 1997