Regulatory

Part:BBa_K342003

Designed by: Nathalie Isorce   Group: iGEM10_INSA-Lyon   (2010-10-24)
Revision as of 09:21, 21 September 2011 by Amateur (Talk | contribs)

OmpR234 protein, with higher effect on Curli promoter


This part is coding for a mutated protein OmpR. This protein will be phosphorylated by the complex EnvZ. This mutated phosphorylated protein is a better activator of the Curli promoter than OmpR.

Usage and Biology

This part can be used to induce a constitutive biofilm producing phenotype in bacteria.


Characterization

Microscopy tests



Quantitative adherence tests in plates



For characterization, the plasmids are introduced in PHL1414 strain. The strain with pIG16 plasmid (18A-OmpR234 in pSB1A2) is noted S31 and the control strain with the pIG3 plasmid (18A part in pSB1A2) is noted S30.

The effect of the plasmid on the adherence is measured.

To that end the strains are seeded in 24 wells plate.

The measure of the OD 600 will give the percentage of adherence of the bacteria and the violet crystal coloration will give us a first idea on the formation of biofilm.


Conclusion: The OD600 seemed to be the same for the two strains. There is no effect of the plasmid on the growth.


The percentage of adherence is now measured for both strains

Conclusion: we can see a strong increase of stickiness when the part 18A-OmpR234 is introduced.



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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 137
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//collections/biofilm
Parameters
None