Difference between revisions of "Part:BBa K1632003"

Revision as of 08:11, 18 September 2015

fim switch[default OFF](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. 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 fim switch(Tokyo_Tech) is similar with fim switch(wild-type).

Fig. 1. Tokyo Tech 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 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
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1102

@@ Line 3: / Line 3: @@
 <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>
-<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.
+<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 <i>fim</i> switch(Tokyo_Tech) is similar with <i>fim</i> switch(wild-type).
 [[Image:Tokyo Tech fim switch Tokyo Tech design.png|thumb|center|600px|Fig. 1. Tokyo Tech ''fim'' switch (Tokyo_Tech/J23119) design (Up:on state  Down:off state)]]<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.
+<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.