Part:BBa_K257011
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
Pbad-clyA-RFP on LB medium (without arabinose)
Pfec-RFP on medium with citrate (without clyA, fluorescence isn't localize on the membrane exclusively)
On the first image, we observed fluorescence localized to the membrane when growing bacteria transformed with PBad-ClyA-RFP on a medium with arabinose. We can compare it to 2 others images from microcoscopy : with Pbad-clyA-RFP without arabinose, the fluorescent is less intense (pBad isn't repressed so there is little expression of clyA-RFP). The third image demonstrate that without clyA, RFP isn't localize exclusively in the membrane (here, it's an example with Bba_K257023)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1302
Illegal BamHI site found at 1144 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
Illegal AgeI site found at 2712
Illegal AgeI site found at 2824 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961
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