Team:GO Paris-Saclay/Contribution
CONTRIBUTION
In the process of reviewing genes that would be good candidates to entangle with another gene, we were looking at new fluorescent proteins. We came across a recent publication that described a thermostable flavin-based fluorescent domain. This domain belonged to a larger protein with histidine kinase activity from the Chloroflexus aggregans, a thermophilic phototrophic bacterium*. The sequence predicted to confer fluorescence, which we called stable_YFP_LOV, was cloned along with a short fragment encoding a tag of 6 histidines. The His-tag could be useful in the future for purification purposes. Using flow cytometry, we showed that upon expression of our new Biobrick in E. coli, bacteria exhibit a green fluorescence. The relatively small size (ca. 11 kDa) of our new part and its described heat stability constitute two attractive features that could be useful for other iGEM teams in the future. More info can be found in the registry.
* Nazarenko et al. 2019, Photochem Photobiol Sci, doi: 10.1039/c9pp00067d
Biobrick part code |
Nickname |
Coding sequence |
Vector used |
BBa K3427000 |
stable_YFP_LOV |
thermostable Yellow Fluorescent Protein |
pBAD24-MoClo |
 |
 |