Difference between revisions of "Part:BBa K3792006"
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Revision as of 01:38, 22 October 2021
NodCIJ Generator I
The NodCIJ Generator I has a BIOFAB promoter with a strength recorded at x fluorescence per cell. The strongest promoter in the collection has a strength of y . NodC is a chitin synthase that produces tetrameric chitooligomers. NodI and NodJ are transport proteins that usually secrete.
This part consists of 2555 base-pair length. It includes a promoter, 3 ribosome binding sites, 3 protein-coding sequences, and a terminator. We used promoter K2832184, a weak promoter from the BIOFAB collection. In addition, we also used protein-coding sequences from Chitin synthase from Rhiozobium leguminosarum, Nodl a protein possibly involved in transporting chitin across the cell membrane, and NodJ, a protein also possibly involved in the transportation of chitin
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.
Cells Supernatant | Supernatant of the Chitin Secretion Cells2.477 |
---|---|
LB and Chitosan Assay | LB Growth Medium with the Chitosan Detection Assay1.515 |
DI Water and Chitosan Assay | Deionized Water with the Chitosan Assay0.084 |
LB and No Assay | LB Growth Medium with No Chitosan Assay0.08 |
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 492
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 854