Difference between revisions of "Part:BBa K1632001"

Revision as of 09:01, 16 September 2015

fim switch[default OFF](Tokyo_Tech/J23119)

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. Tokyo Tech fim switch (Tokyo_Tech/J23119) design). So the promoter can easily be interchanged. Except for insertion of restriction enzyme sites, basically, the design of TokyoTech fim switch is similar with Wild Type fim switch.

Fig. 1. Tokyo Tech fim switch (Tokyo_Tech/J23119) design

We constructed fim switch [default OFF](Tokyo_Tech/J23119)_rbs_gfp(BBa_K1632003) to characterize the function of this part, by inserting this promoter upstream of a GFP coding sequence.
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 98
Illegal NheI site found at 121
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 92
Illegal BamHI site found at 133
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1632001 short</partinfo>
-The <i>fim</i> 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 <i>fim</i> switch by adding the Fim recombinase.<br>
+<span style="margin-left: 10px;">The <i>fim</i> 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 <i>fim</i> switch by adding the Fim recombinase.<br>
-This fim switch has a J23119 promoter. Also between the promoter and the inverting site, there are restriction enzyme sites. Therefore, the promoter can easily be interchanged.
+<span style="margin-left: 10px;">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. Tokyo Tech ''fim'' switch (Tokyo_Tech/J23119) design). So the promoter can easily be interchanged. Except for insertion of restriction enzyme sites, basically, the design of TokyoTech fim switch is similar with Wild Type fim switch.
+[[Image:Tokyo Tech fim switch Tokyo Tech design.png|thumb|center|600px|Fig. 1. Tokyo Tech ''fim'' switch (Tokyo_Tech/J23119) design]]<br>
+<span style="margin-left: 10px;">We constructed ''fim'' switch [default OFF](Tokyo_Tech/J23119)_rbs_gfp(<partinfo>BBa_K1632003</partinfo>) to characterize the function of this part, by inserting this promoter upstream of a GFP coding sequence. <br>
+<span style="margin-left: 10px;">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.