Team:Toulouse/Parts
NE PAS MODIFIER PROUT
In the list below you will find an overview over the BioBrick parts we added to the registry, which were created by the iGEM Toulouse 2015 team.
For our project, we worked on three different modules : attract the varroa, kill the mite and finally the circadian switch to alternatively produce the two molecules of interest, butyrate during the day and formate during the night.
| Name | Type | Genic construction | Lenghth (bp) | References |
|---|---|---|---|---|
| BBa_K1587000 | Composite (with RBS) | RBS-ho1 | 744 | [13] |
| BBa_K1587001 | Basic part | tesB | 861 | [3][4][5] |
| BBa_K1587002 | Composite (with RBS) | RBS-pcyA | 796 | [13] |
| BBa_K1587003 | Basic part | crt | 786 | [1] [2] [3] [4] [7] |
| BBa_K1587004 | Basic part (others) | pBla-RBS-ccr-RBS-hbd-RBS-crt-RBS- tesB-RBS-atoB-Terminator | 5175 | [1] [2] [3] [4] [6] |
| BBa_K1587005 | Basic part (others) | pBla-RBS-hbd-RBS-crt-RBS- tesB-RBS-atoB-Terminator | 3827 | [1] [2] [3] [4] [6] |
| BBa_K1587006 | Basic part (others) | pOmpC-LacIbox-RBS-cI | 905 | [12][13] |
| BBa_K1587007 | Basic part (others) | RBS-pflB-RBS-pflA-Terminator | 3093 | [7] [8] [9] [10] [11] |
Attraction (butyrate pathway)
The chassis we used is Escherichia coli, and this bacterium is not able to naturally produce butyrate. That is why we introduced genes from others bacterial strains to synthesize this molecule.
Basic parts
tesB (BBa_K1587001)
Gene from Escherichia coli involved in the butyrate pathway that enables its production directly from acyl-coAs. This group of enzymes catalyzes the hydrolysis of acyl-CoAs into free fatty acid (in our case, butyryl-coA into butyrate) plus reduced coenzyme A (CoA-SH).
crt (BBa_K1587003)
Gene from Clostridium acetobutylicum was introduced in our bacterium after codon optimization in order to obtain a better expression in E. coli. The crt enzyme substrate is 3-hydroxybutyryl CoA, and the product is Crotonyl CoA. This reaction does not need any coenzyme.
Other parts
ccr-Butyrate pathway (BBa_K1587004)
This BioBrick construction is composed of a constitutive promoter p(Bla) (BBa_I14018) and 5 genes from three different micro-organisms : in yellow are the E.coli genes, in blue those from Clostridium acetobutylicum and finally, the purple gene is from Streptomyces collinus. A Ribosome Binding Site (RBS) represented by a green circle (BBa_B0030), is added between each gene in order to improve the proteic synthesis. Finally, a strong terminator (BBa_B1006) represents the end of the sequence.
- tesB and crt have been described previously.
- ccr encodes crotonyl CoA reductase, an oxidoreductase which acts on the double bond CH=CH. hbd in Clostridium acetobutylicum encodes 3-hydroxybutyryl-CoA dehydrogenase, an oxidoreductase which catalyses the formation of alcohol function. atoB, in E.coli, encodes acetyl CoA acetyltransferase which catalyses the condensation of two acetyl CoA.
