Difference between revisions of "Part:BBa K1632002"
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<partinfo>BBa_K1632002 parameters</partinfo>
 
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===References===
 
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[1]Ian C. Blomfield <i>et al</i>. (1997) Integration host factor stimulates both FimB- andFimE-mediated site-specific DNA inversion that controlsphase variation of type 1 fimbriae expression in Escherichia coli. Molecular Microbiology 23(4), 705–717
 
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[2]John M. Abraham <i>et al</i>. (1985) An invertible element of DNA controls phase variation of type 1 fimbriae of Escherichia coli. Proc Natl Acad Sci U S A 82(17):5724-7
 
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[3]Matthew P. McCusker <i>et al</i>. (2008) DNA sequence heterogeneity in Fim tyrosine-integrase recombinase-binding elements and functional motif asymmetries determine the directionality of the fim genetic switch in Escherichia coli K-12. Molecular Microbiology 67(1): 171–187
 

Revision as of 09:47, 18 September 2015

fim switch[default ON](Tokyo_Tech/J23119)_rbs_gfp

The fim switch is the promoter containing repeated DNA sequence which is inverted by the Fim recombinase. Therefore, we can control the expression of the gene downstream of the fim switch by adding the Fim recombinase.
We designed this fim switch which has a J23119 promoter. Also between the promoter and the inverting site, there are two restriction enzyme sites in each front (SalIand BamHI) and back (BglII and MluI)(Fig. 1. fim switch (Tokyo_Tech/J23119) design). So the promoter can easily be interchanged. Except for insertion of restriction enzyme sites, basically, the design of fim switch(Tokyo_Tech) is similar with fim switch(wild-type).

Fig. 1. fim switch (Tokyo_Tech/J23119) design (Up:on state Down:off state)

From our results of our assay, the inversion of fim switch (Tokyo_Tech/J23119) by FimB/FimE was not confirmed correctly. The FimB protein inverts fim switch in the ON-to-OFF and the OFF-to-ON direction correctly. However the FimE protein didn’t inverts fim switch predominantly in the ON-to-OFF direction. In the assay, the FimE protein inverts fim switch in the ON-to-OFF and the OFF-to-ON direction. In other words, the FimE protein works as the FimB protein.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 345
    Illegal NheI site found at 368
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 374
    Illegal BamHI site found at 333
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1102