Difference between revisions of "Team:Paris Bettencourt/Project/VitaminB2"
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<p>Genetic sequences of <i>Bacillus subtilis</i> four enzymes were codon optimized for <i>Lactobacillus plantarum</i> on IDT website tool.<br> | <p>Genetic sequences of <i>Bacillus subtilis</i> four enzymes were codon optimized for <i>Lactobacillus plantarum</i> on IDT website tool.<br> | ||
− | The enzyme was expressed by a promoter rather than a single operon expression to allow a precise tuning of the gene transcription. Two bottlenecks were identified in the pathway (M. Birkenmeier, Biotech. Lett., 2014). The first bottleneck correspond to the GTP cyclohydrolase activity of RibA. Overcoming the first bottleneck creates a second bottleneck corresponding to RibT's lumazine synthase activity.<br> | + | The enzyme was expressed by a promoter rather than a single operon expression to allow a precise tuning of the gene transcription. Two metabolic bottlenecks were identified in the pathway (M. Birkenmeier, Biotech. Lett., 2014). The first bottleneck correspond to the GTP cyclohydrolase activity of RibA. Overcoming the first bottleneck creates a second bottleneck corresponding to RibT's lumazine synthase activity.<br> |
To promote a differential expression of the four genes, we used the two synthetic <i>Lactobacillus</i> promoters p25 and p48 (respectively medium and strong expressing promoters) (I. Rud, Microbiology, 2006).<br> | To promote a differential expression of the four genes, we used the two synthetic <i>Lactobacillus</i> promoters p25 and p48 (respectively medium and strong expressing promoters) (I. Rud, Microbiology, 2006).<br> | ||
For translation initiation, we used the consensus RBS for <i>Lactobacillus</i> (Tauer et al. Microbial Cell Factories 2014, 13:150).<br> | For translation initiation, we used the consensus RBS for <i>Lactobacillus</i> (Tauer et al. Microbial Cell Factories 2014, 13:150).<br> |
Revision as of 15:01, 18 September 2015