Difference between revisions of "Part:BBa K3792010"
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<partinfo>BBa_K3792010 short</partinfo> | <partinfo>BBa_K3792010 short</partinfo> | ||
− | The NodC Control Generator with Medium Promoter produces a medium quantity of the NodC protein. The NodC protein | + | The NodC Control Generator with Medium Promoter produces a medium quantity of the NodC protein. The NodC protein produces the carbohydrate, chitin. The NodC Generator with Weak Promoter does not produce a lot of the NodC protein. The NodC protein produces chitin. |
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+ | This part consists of 832 base-pair length. It includes a promoter, 1 ribosome binding site, 1 protein-coding sequence, and a terminator. We used promoter K2832184, a weak promoter from the BIOFAB collection. In addition, we also used a protein-coding sequence from Chitin synthase from Rhiozobium leguminosarum. | ||
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+ | This generator was assembled into the pSB1K3 plasmid, and then transformed into E.Coli NEB 5-Alpha. To test if the cells were secreting chitin, The FSU 2021 team used the Chitosan Assay Kit (XAN-5126) available from Cell Biolabs. The manual for this kit contains the protocols for converting chitin to chitosan, along with a protocol for detecting chitosan. | ||
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+ | Chitin from the cells was converted into chitosan by reacting with 12.5M NaOH at 95°C for 24 hours. The solution was then neutralized with acetic acid. The assay was then performed on the supernatant of the cell culture. | ||
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Revision as of 01:06, 20 October 2021
NodC Generator II
The NodC Control Generator with Medium Promoter produces a medium quantity of the NodC protein. The NodC protein produces the carbohydrate, chitin. The NodC Generator with Weak Promoter does not produce a lot of the NodC protein. The NodC protein produces chitin.
This part consists of 832 base-pair length. It includes a promoter, 1 ribosome binding site, 1 protein-coding sequence, and a terminator. We used promoter K2832184, a weak promoter from the BIOFAB collection. In addition, we also used a protein-coding sequence from Chitin synthase from Rhiozobium leguminosarum.
This generator was assembled into the pSB1K3 plasmid, and then transformed into E.Coli NEB 5-Alpha. To test if the cells were secreting chitin, The FSU 2021 team used the Chitosan Assay Kit (XAN-5126) available from Cell Biolabs. The manual for this kit contains the protocols for converting chitin to chitosan, along with a protocol for detecting chitosan.
Chitin from the cells was converted into chitosan by reacting with 12.5M NaOH at 95°C for 24 hours. The solution was then neutralized with acetic acid. The assay was then performed on the supernatant of the cell culture.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 503
- 1000COMPATIBLE WITH RFC[1000]