Difference between revisions of "Part:BBa K2971001"
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<partinfo>BBa_K2971001 short</partinfo> | <partinfo>BBa_K2971001 short</partinfo> | ||
| − | + | This part is the gene crtI (dr0861) from the extremophile Deinococcus radiodurans and encodes | |
| − | This part is the gene | + | phytoene desaturase (CrtI). CrtI catalyzes the desaturation of phytoene (a colorless compound) into |
| − | + | lycopene (a deep red compound)[1]. The desaturation produces a polyene chromophore. It is this | |
| + | chromophore that is utilized in the biogenic solar cell (see design page) to absorb energy from light. | ||
| − | + | The UiOslo 2019 team used this gene in a composite part (BBa_K2971004) to produce the red pigment lycopene, in | |
| − | The UiOslo 2019 team used this gene in a composite part to produce the red pigment lycopene, in Escherichia coli, by | + | <i>Escherichia coli</i>, by inserting it into a vector along with the genes <i>crtE</i> and <i>crtI</i>. This composite part produced red cells when expressed in Escherichia coli confirming that the part functions as intended. |
| − | </ | + | |
| + | The part was cloned into an expression vector under the arabionose inducible promotor <i>araC</i>. Successful cloning was confirmed with colony PCR of transformed colonies (figure 1). Colonies that carried the insert was sequenced to check for the potential presence of mutations or frame-shifts. | ||
| + | |||
| + | We expressed the protein in <i>E. coli</i> (DH5α)(figure 2). We detected a faint protein band during our SDS analysis; it is likely the expression was not well-optimized. | ||
'''References''' | '''References''' | ||
Revision as of 22:44, 21 October 2019
crtI from Deinococcus radiodurans
This part is the gene crtI (dr0861) from the extremophile Deinococcus radiodurans and encodes phytoene desaturase (CrtI). CrtI catalyzes the desaturation of phytoene (a colorless compound) into lycopene (a deep red compound)[1]. The desaturation produces a polyene chromophore. It is this chromophore that is utilized in the biogenic solar cell (see design page) to absorb energy from light.
The UiOslo 2019 team used this gene in a composite part (BBa_K2971004) to produce the red pigment lycopene, in Escherichia coli, by inserting it into a vector along with the genes crtE and crtI. This composite part produced red cells when expressed in Escherichia coli confirming that the part functions as intended.
The part was cloned into an expression vector under the arabionose inducible promotor araC. Successful cloning was confirmed with colony PCR of transformed colonies (figure 1). Colonies that carried the insert was sequenced to check for the potential presence of mutations or frame-shifts.
We expressed the protein in E. coli (DH5α)(figure 2). We detected a faint protein band during our SDS analysis; it is likely the expression was not well-optimized.
References
1. Schaub P, Yu Q, Gemmecker S, Poussin-Courmontagne P, Mailliot J, McEwen AG, et al. (2012) On the Structure and Function of the Phytoene Desaturase CRTI from Pantoea ananatis, a Membrane-Peripheral and FAD-Dependent Oxidase/Isomerase. PLoS ONE 7(6): e39550. https://doi.org/10.1371/journal.pone.0039550
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 955
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]