Difference between revisions of "Part:BBa K3792006"
Anishshahez (Talk | contribs) |
|||
Line 1: | Line 1: | ||
+ | {{FSU/ChartsCSS}} | ||
+ | |||
+ | <html> | ||
+ | <style> | ||
+ | #modular-promoter-library-chart.column { | ||
+ | height: 400px; | ||
+ | max-width: 100% !important; | ||
+ | margin: 0 auto; | ||
+ | --labels-size: 0.5rem; | ||
+ | } | ||
+ | |||
+ | #modular-promoter-library-chart tr { | ||
+ | transition-duration: 0.3s; | ||
+ | } | ||
+ | #modular-promoter-library-chart tr:hover { | ||
+ | background-color: rgba(0, 0, 0, 0.2); | ||
+ | } | ||
+ | #modular-promoter-library-chart tr:hover th { | ||
+ | background-color: rgba(0, 0, 0, 0.4); | ||
+ | color: #fff; | ||
+ | } | ||
+ | </style> | ||
+ | </html> | ||
+ | |||
__NOTOC__ | __NOTOC__ |
Revision as of 00:41, 22 October 2021
NodCIJ Generator I
The NodCIJ Generator with a weak promoter does not produce a lot of the NodC protein, NodI protein, and NodJ protein. The NodC protein produces chitin and NodI and NodJ transports nodulation factors.
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.
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