Difference between revisions of "Part:BBa K2992028"

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(→‎Usage and Biology)
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===Usage and Biology===
 
===Usage and Biology===
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This parts entry represents a promoterless acetone-production pathway for plasmid-borne expression in <i>C. sporogenes</i>. 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_M36587 BBa_M36587] coupled with the two units of the ctfAB 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]. 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 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_M36587 BBa_M36587] 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 [] and associated 5’UTR+RBS [] from the non-toxic non-haemagglutinin gene of C. botulinum ntnH. 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>
  
 
===Characterisation===
 
===Characterisation===

Revision as of 08:05, 25 September 2019


Acetone pathway: ca_thl -cb_ctfAB-ca_adc-cpa_Tfdx

Promoterless acetone production pathway with C. acetobutylicum thl and adc with C. botulinum ctfAB

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_M36587 coupled with the two units of the ctfAB>/i> complex from <i>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 [] and associated 5’UTR+RBS [] from the non-toxic non-haemagglutinin gene of C. botulinum ntnH. 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 355
    Illegal PstI site found at 3250
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 3250
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 355
    Illegal PstI site found at 3250
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 355
    Illegal PstI site found at 3250
  • 1000
    COMPATIBLE WITH RFC[1000]

References

Heap Modular Cornillo et al 1997