Part:BBa_K2255004
Xylella fastidiosa constitutive promoter + strong RBS
This sequence is a strong constitutive promoter with a strong RBS designed for Xylella fastidiosa.
It is the upstream sequence of a putative highly transcribed gene from Xylella fastidiosa 9a5c, found by analyzing the Best Bidirectional Hits (BBH) between Escherichia coli str. K-12 substr. MG1655 genes and Xylella fastidiosa 9a5c.
Usage and Biology
This part enable us to produce in high quantities proteins in Xylella fastidiosa. It was engineered for the phage-like particle design of our project. Check out our [http://2017.igem.org/Team:Aix-Marseille/M13_Design design page] for more informations.
To evaluate the activity of the promoter and RBS, sfGFP protein was expressed in Escherichia coli DH5α bacteria and it fluorescence properties was studied. We compared our promoter and RBS to a well studied biobrick BBa_K608002 which is an E. coli strong promoter and RBS.
We used [http://2017.igem.org/Competition/InterLab_Study/Plate_Reader InterLab protocol] and negative control to test the fluorescence properties of these promoters and RBS.
We can observe that bacteria transformed with the plasmid containing BBa_K608002 gives a increase of fluorescence with time that those with X. fastidiosa RBS and promoter (BBa_K2255004) doesn't have. Moreover, it seems that bacteria transformed with the plasmid containing BBa_K2255004 tend to stay at a basal level along with the negative control.
Thus we can assume that X. fastidiosa RBS and promoter is not functionnal in E. coli DH5α bacteria, but as this part was design to be a promoter and RBS for X. fastidiosa we must test the fluorence activity in this bacteria in oreder to have a proper comparaison.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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