Difference between revisions of "Part:BBa K3738028"

Latest revision as of 02:07, 22 October 2021

MlrA N-terminal 6XHistidine Tag and C-Terminal and Anionic Tag (Basic)

MlrA, or microcystinase, cleaves the peptide backbone of MC-LR cyanotoxins produced by Microcystis aeruginosa cyanobacteria. This crucial step results in the peptide losing the cyclic structure, and therefore the majority of its toxicity. Further degradation continues in an enzymatic pathway by interactions with other peptidolytic enzymes from the microcystinase gene cluster. This part originally comes from the Aalto-Helsinki 2016 iGEM team (Part BBa_K1907001), but was originally codon-optimized for yeast. We have modified this part by codon-optimizing it for use in E.coli. MlrA is known to be taken up into the periplasm with aid of transport protein MlrD (Saito et al., 2003), however for the mlrA sequence from Sphingomonas sp. USTB-05 the mlrD sequence is currently unavailable. Thus we have added both a histidine tag and anionic tag for purification and MS2-encapsulation respectively which will enable it to be packaged and introduced via MS2 into cyanobacteria

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 184
Illegal BglII site found at 925
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 867

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 <partinfo>BBa_K3738028 short</partinfo>
-MlrA, or microcystinase, cleaves the peptide backbone of MC-LR cyanotoxins produced by <i>Microcystis aeruginosa</i> cyanobacteria. This crucial step results in the peptide losing the cyclic structure, and therefore the majority of its toxicity. Further degradation continues in an enzymatic pathway by interactions with other peptidolytic enzymes from the microcystinase gene cluster. This part originally comes from the Aalto-Helsinki 2016 iGEM team (Part BBa_K1907001), but was originally codon-optimized for yeast. We have modified this part by codon-optimizing it for use in E.coli. MlrA is known to be taken up into the periplasm with aid of transport protein MlrD (Saito et al., 2003), however for the mlrA sequence from <i>Sphingopyxis sp.</i> USTB-05 the mlrD sequence is currently unavailable. Thus we have added both a histidine tag and anionic tag for purification and MS2-encapsulation respectively which will enable it to be packaged and introduced via MS2 into cyanobacteria
+MlrA, or microcystinase, cleaves the peptide backbone of MC-LR cyanotoxins produced by <i>Microcystis aeruginosa</i> cyanobacteria. This crucial step results in the peptide losing the cyclic structure, and therefore the majority of its toxicity. Further degradation continues in an enzymatic pathway by interactions with other peptidolytic enzymes from the microcystinase gene cluster. This part originally comes from the Aalto-Helsinki 2016 iGEM team (Part BBa_K1907001), but was originally codon-optimized for yeast. We have modified this part by codon-optimizing it for use in E.coli. MlrA is known to be taken up into the periplasm with aid of transport protein MlrD (Saito et al., 2003), however for the mlrA sequence from <i>Sphingomonas sp.</i> USTB-05 the mlrD sequence is currently unavailable. Thus we have added both a histidine tag and anionic tag for purification and MS2-encapsulation respectively which will enable it to be packaged and introduced via MS2 into cyanobacteria
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