Difference between revisions of "Part:BBa K2992036"

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 [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>.This operon is regulated by the promoter [https://parts.igem.org/Part:BBa_K2992001 BBa_K2992001] and associated 5’UTR+RBS [https://parts.igem.org/Part:BBa_K2992015 BBa_K2992015] from the non-toxic non-haemagglutinin <i>ntnH</i> gene of <i>C. botulinum</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>