Team:Aix-Marseille/Basic Part
We made three basic parts during this years iGEM competition coding for 2 different enzymes.
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BBa_K2718005
Methionine gamma-lyase (MGL) catalyzes the conversion of l-methionine into methanthiol, then the oxidation of methanethiol produces DMDS (Dimethyl disulfide) and DMTS ( Dimethyl trisulfide). BBa_K2718005 is an RFC 10 compatible part that we derived from BBa_K1493300 using PCR to add a his tag. To test activity we used the compositive part BBa_K2718006
BBa_K2718020
The Chit1 gene that codes for the endochitinase from the fungus Beauveria bassiana, an enzyme that degrads chitin, was made as BBa_K2718020. It's endochitinase wich hydrolyses 1-4 beta link between N-acetyl-D-glucosamine monomeres the sequence comes from uniprot, we used the protein sequence between amino acid 41 to 330, that corresponds to the catalitic domain. We improved the DNA sequence with the IDT tool to optimize it for synthesis by E.coli. Part used the RFC 25 standard. We tested the activiy using Schales' method[1] with the part BBa_K2718022
BBa_K2718021
Chitinase BBa_K2718020is a fungus (Beauveria bassiana) enzyme that degrads chitin. It's endochitinase wich hydrolyses 1-4 beta link between N-acetyl-D-glucosamine monomere . Sequence comes from uniprot, we used peptidic sequence between 41 to 330 AA, that's correspond to catalitic domain. We improve nucleotidic sequence with IDT tool for synthesis by E.coli, and there is his tag in C-ter. Part is RFC 25 standards. We test activiy with Schales' procedure[2].