Composite
PBad-clyA-

Part:BBa_K257011

Designed by: Soufiane Boumahdi   Group: iGEM09_Paris   (2009-10-17)
Revision as of 00:18, 22 October 2009 by Sboumahdi (Talk | contribs) (→‎Experience)

PBad + clyA_CTerm + RFP_NTerm

This part allows us to express the fusion protein clyA-RFP. In a mutant strain which constitutively produce vesicles, the fusion protein inserted in the outer membrane of the bacteria will be incorporated, during vesiculation, into the lumen of the vesicles. The promoter pBad is inductible with L-arabinose.


Experience

Pbad-clyA-RFP on arabinose medium

TolA clyA Ara 01.jpg


Pbad-clyA-RFP on LB medium (without arabinose)

TolA clyA LB 01.jpg

Pfec-RFP on medium with citrate (without clyA, fluorescence isn't localize on the membrane exclusively)


File:Citrate 30.jpg

We observed fluorescence localized to the membrane when growing bacteria transformed with PBad-ClyA-RFP on a medium with arabinose.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1302
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
    Illegal AgeI site found at 2712
    Illegal AgeI site found at 2824
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


[edit]
Categories
Parameters
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