Difference between revisions of "Team:Aix-Marseille/Basic Part"
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{{Aix-Marseille/top|title=Basic Parts}} | {{Aix-Marseille/top|title=Basic Parts}} | ||
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| + | We made three basic parts during this years iGEM competition coding for 2 different enzymes. | ||
| + | <!-- Pour chaque part il serait bien d'expliquer d'ou vien la sequence (avec une reference), ce que c'est censé faire, comment on a testé ce que ca fait --> | ||
| + | ==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). [http://parts.igem.org/Part:BBa_K2718005 BBa_K2718005] is an RFC 10 compatible part that we derived from [http://parts.igem.org/Part:BBa_K1493300 BBa_K1493300] using PCR to add a his tag. To test activity we used the compositive part [http://parts.igem.org/Part:BBa_K2718006 BBa_K2718006] | ||
| + | [https://static.igem.org/mediawiki/parts/1/16/--Aix-Marseille--Shcema_MGL_parts.jpg] | ||
| − | + | ==BBa_K2718020== | |
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| − | + | The Chit1 gene that codes for the endochitinase from the fungus ''Beauveria bassiana'', an enzyme that degrads chitin, was made as [http://parts.igem.org/Part:BBa_K2718020 BBa_K2718020]. It's endochitinase wich hydrolyses 1-4 beta link between N-acetyl-D-glucosamine monomeres the sequence comes from [https://www.uniprot.org/uniprot/Q8J1Y3 uniprot], we used the protein sequence between amino acid 41 to 330, that corresponds to the [https://www.ebi.ac.uk/interpro/protein/Q8J1Y3 catalitic domain]. We improved the DNA sequence with the [https://eu.idtdna.com/CodonOpt IDT tool] to optimize it for synthesis by ''E.coli''. Part used the RFC 25 standard. We tested the activiy using Schales' method[https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3975300/pdf/1754-6834-7-37.pdf] with the part | |
| − | + | [http://parts.igem.org/Part:BBa_K2718022 BBa_K2718022] | |
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| + | ==BBa_K2718021== | ||
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| + | This part is the same as the chitinase [http://parts.igem.org/Part:BBa_K2718020 BBa_K2718020] from ''Beauveria bassiana'' but we added a C-terminal oligo-histidine tag to the protein to facilitate purification and detection using a western blot. The part used the RFC 25 standard. We test enzyme activity of this part, and the availability of the his-tag for purification as [http://parts.igem.org/Part:BBa_K2718020 BBa_K2718020]. | ||
Latest revision as of 01:28, 18 October 2018
Basic Parts
We made three basic parts during this years iGEM competition coding for 2 different enzymes.
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 [1]
![[1]](https://static.igem.org/mediawiki/parts/1/16/--Aix-Marseille--Shcema_MGL_parts.jpg){kind=link}
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[2] with the part BBa_K2718022
BBa_K2718021
This part is the same as the chitinase BBa_K2718020 from Beauveria bassiana but we added a C-terminal oligo-histidine tag to the protein to facilitate purification and detection using a western blot. The part used the RFC 25 standard. We test enzyme activity of this part, and the availability of the his-tag for purification as BBa_K2718020.