Difference between revisions of "Part:BBa K2992034"
(→Usage and Biology) |
|||
Line 6: | Line 6: | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
− | 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 | + | 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 [https://parts.igem.org/Part:BBa_M36585 BBa_M36585] coupled with the two units of the <i>ctfAB</i> complex [https://parts.igem.org/Part:BBa_ M36581 BBa_ M36581] and [https://parts.igem.org/Part:BBa_ M36582 BBa_ M36582] which are all derived from <i>C. acetobutylicum</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> |
<!-- --> | <!-- --> | ||
Revision as of 11:27, 26 September 2019
Acetone pathway: ca_thl-ca_ctfAB-cp_TFdx
Promoterless acetone production pathway with C. acetobutylicum thl, ctfAB and adc genes.
Usage and Biology
This parts entry represents a promoterless acetone-production pathway for plasmid-borne expression in C. sporogenes. The entry comprises the thiolase gene thl BBa_K2992008 and acetoacetate decarboxylase adc gene BBa_M36585 coupled with the two units of the ctfAB complex M36581 BBa_ M36581 and M36582 BBa_ M36582 which are all derived from C. acetobutylicum. 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
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 355
Illegal PstI site found at 1894
Illegal PstI site found at 3159 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1894
Illegal PstI site found at 3159 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 355
Illegal PstI site found at 1894
Illegal PstI site found at 3159 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 355
Illegal PstI site found at 1894
Illegal PstI site found at 3159 - 1000COMPATIBLE WITH RFC[1000]
References
Heap Modular Cornillo et al 1997