Difference between revisions of "Team:Aix-Marseille/Composite Part"
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Methionine gamma-lyase (MGL) catalyzes the conversion of l-methionine to methanethiol, the oxidation of methanethiol then produces first DMDS (Dimethyl disulfide) and then DMTS ( Dimethyl trisulfide). The composite part [http://parts.igem.org/Part:BBa_K2718006 BBa_K2718006] is made from [http://parts.igem.org/Part:BBa_K2718005 BBa_K2718005], an RFC 10 compatible part that we derived from [http://parts.igem.org/Part:BBa_K1493300 BBa_K1493300] using a PCR to add a C-terminal histidine tag, preceded by the [http://parts.igem.org/wiki/index.php?title=Part:BBa_R0011 BBa_R0011], an IPTG inducible promoter part. | Methionine gamma-lyase (MGL) catalyzes the conversion of l-methionine to methanethiol, the oxidation of methanethiol then produces first DMDS (Dimethyl disulfide) and then DMTS ( Dimethyl trisulfide). The composite part [http://parts.igem.org/Part:BBa_K2718006 BBa_K2718006] is made from [http://parts.igem.org/Part:BBa_K2718005 BBa_K2718005], an RFC 10 compatible part that we derived from [http://parts.igem.org/Part:BBa_K1493300 BBa_K1493300] using a PCR to add a C-terminal histidine tag, preceded by the [http://parts.igem.org/wiki/index.php?title=Part:BBa_R0011 BBa_R0011], an IPTG inducible promoter part. | ||
To test the activity of [http://parts.igem.org/Part:BBa_K2718006 BBa_K2718006] we produced and purified the his-tagged protein and used DTNB to measure the enzyme activity. DTNB reacts with the released thiol group [https://www.dovepress.com/activities-of-methionine-gamma-lyase-in-the-acidophilic-archaeon-ldquo-peer-reviewed-article-RRB]. | To test the activity of [http://parts.igem.org/Part:BBa_K2718006 BBa_K2718006] we produced and purified the his-tagged protein and used DTNB to measure the enzyme activity. DTNB reacts with the released thiol group [https://www.dovepress.com/activities-of-methionine-gamma-lyase-in-the-acidophilic-archaeon-ldquo-peer-reviewed-article-RRB]. | ||
Revision as of 17:02, 17 October 2018
Composite Parts
We constructed 4 composite parts during this years iGEM competition. Three of them were made by attaching a promoter (with or without a ribosome binding site) in front of another part. The last was an RBS coding sequence fusion that we had synthesised by IDT.
BBa_K2718006
Methionine gamma-lyase (MGL) catalyzes the conversion of l-methionine to methanethiol, the oxidation of methanethiol then produces first DMDS (Dimethyl disulfide) and then DMTS ( Dimethyl trisulfide). The composite part BBa_K2718006 is made from BBa_K2718005, an RFC 10 compatible part that we derived from BBa_K1493300 using a PCR to add a C-terminal histidine tag, preceded by the BBa_R0011, an IPTG inducible promoter part. To test the activity of BBa_K2718006 we produced and purified the his-tagged protein and used DTNB to measure the enzyme activity. DTNB reacts with the released thiol group [1].
BBa_K2718010
BBa_K2718011
BBa_K2718022
Our part BBa_K2718022 is made by fusing the promoter RBS in part BBa_J04500 to our chitinase part with a C-terminal oligo-histidine tag BBa_K2718021. We tested the enzyme activity using Schales' method[2] we also tested production of the protein and used the his-tag to purify the protein.