Difference between revisions of "Part:BBa K3229301"

Revision as of 15:47, 21 October 2019

McyD promoter reporter

This part is appropriate to measure the activity level of the putative mcyD promoter region. Normally we can find protein genes (McyABC and McyDEFGHIJ) next to the promoter in both 5'-3' and 3'-5' directions, thus the promoter can regulate the formation of the microcystin toxin through the synthesis of microcystin producing proteins. When the toxin is present it enhances the transcription from the promoter due to the phenomenon called autoinduction. We replaced the toxin-producing protein genes with GFP, thus when microcystin appears it induces the production of GFP. We will observe the emitted light with widely used fluorescence detector whether the emitted light is correlated with the amount of toxin. This part consists of the McyD promoter, RBS, GFP and a terminator sequence. We would like to transform our plasmid into Microcystis aeruginosa apart from E. coli. A shuttle plasmid that can replicate both in E. coli and Microcystis is required for this procedure, therefore we put a non-coding ATATATATAT sequence and a BamHI restriction site at the end of our construct. This enables us to insert the construct into the shuttle plasmid.

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 578
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1565

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 <partinfo>BBa_K3229301 short</partinfo>
-This part is appropriate to measure the activity level of the putative mcyD promoter region. Normally we can find protein genes (McyABC and McyDEFGHIJ) next to the promoter in both 5'-3' and 3'-5' directions, thus the promoter can regulate the formation of the microcystin toxin through the synthesis of microcystin producing proteins. When the toxin is present it enhances the transcription from the promoter due to the phenomenon called autoinduction. We replaced the toxin-producing protein genes with GFP, thus when microcystin appears it induces the production of GFP. We will observe with fluorescence microscope whether the emitted light is correlated with the amount of toxin. This part consists of the McyD promoter, RBS, GFP and a terminator sequence. We would like to transform our plasmid into Microcystis aeruginosa apart from E. coli. A shuttle plasmid that can replicate both in E. coli and Microcystis is required for this procedure, therefore we put a non-coding ATATATATAT sequence and a BamHI restriction site at the end of our construct. This enables us to insert the construct into the shuttle plasmid.
+This part is appropriate to measure the activity level of the putative mcyD promoter region. Normally we can find protein genes (McyABC and McyDEFGHIJ) next to the promoter in both 5'-3' and 3'-5' directions, thus the promoter can regulate the formation of the microcystin toxin through the synthesis of microcystin producing proteins. When the toxin is present it enhances the transcription from the promoter due to the phenomenon called autoinduction. We replaced the toxin-producing protein genes with GFP, thus when microcystin appears it induces the production of GFP. We will observe the emitted light with  widely used fluorescence detector  whether the emitted light is correlated with the amount of toxin. This part consists of the McyD promoter, RBS, GFP and a terminator sequence. We would like to transform our plasmid into Microcystis aeruginosa apart from E. coli. A shuttle plasmid that can replicate both in E. coli and Microcystis is required for this procedure, therefore we put a non-coding ATATATATAT sequence and a BamHI restriction site at the end of our construct. This enables us to insert the construct into the shuttle plasmid.
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