Difference between revisions of "Part:BBa K2255004"

Latest revision as of 23:01, 14 October 2017

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

Fluorence levels of Escherichia coli DH5α bacteria transformed with pSB1C3 that contain sfGFP combined with various promoter and RBS.

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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 8: / Line 8: @@
 ===Usage and Biology===
+[[File:T--Aix-Marseille--FluoXpr.png|400px|right|thumb|Fluorence levels of ''Escherichia coli'' DH5α bacteria transformed with pSB1C3 that contain [https://parts.igem.org/Part:BBa_K1321337 sfGFP] combined with various promoter and RBS.]]
 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.
@@ Line 14: / Line 17: @@
 We used [http://2017.igem.org/Competition/InterLab_Study/Plate_Reader InterLab protocol] and [https://parts.igem.org/Part:BBa_R0040 negative control] to test the fluorescence properties of these promoters and RBS.
+We can observe that bacteria transformed with the plasmid containing [https://parts.igem.org/Part:BBa_K608002 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.
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