Difference between revisions of "Part:BBa K1632006"
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<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;">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 the ''fim'' switch which has a J23119 promoter(<partinfo>BBa_K1632000</partinfo>) . Basically, the design of the ''fim'' switch(Tokyo_Tech) is similar to the ''fim'' switch(wild-type). The only difference is that we inserted restriction cut enzyme sites to fim switch(Tokyo_Tech). In detail, in ''fim'' switch(Tokyo_Tech) the sigma 70 promoter is exchanged to the J23119 promoter and there are two restriction enzyme cut sites inserted each in the front (SalI and BamHI) and in the back (BglII and MluI) of the promoter. Due to the insertion of the restriction enzyme cut sites, ''fim'' switch(Tokyo_Tech) has a promoter which can we exchanged with an arbitrary promoter. We actually changed J23119 promoter in the ''fim'' switch(Tokyo_Tech) to Lac promoter(<partinfo>BBa_B0010</partinfo>) as shown in the figure below. | + | <span style="margin-left: 10px;">We designed the ''fim'' switch which has a J23119 promoter(<partinfo>BBa_K1632000</partinfo>) . Basically, the design of the ''fim'' switch(Tokyo_Tech) is similar to the ''fim'' switch(wild-type). The only difference is that we inserted restriction cut enzyme sites to ''fim'' switch(Tokyo_Tech). In detail, in ''fim'' switch(Tokyo_Tech) the sigma 70 promoter is exchanged to the J23119 promoter and there are two restriction enzyme cut sites inserted each in the front (SalI and BamHI) and in the back (BglII and MluI) of the promoter. Due to the insertion of the restriction enzyme cut sites, ''fim'' switch(Tokyo_Tech) has a promoter which can we exchanged with an arbitrary promoter. We actually changed J23119 promoter in the ''fim'' switch(Tokyo_Tech) to Lac promoter(<partinfo>BBa_B0010</partinfo>) as shown in the figure below. |
[[Image:Tokyo Tech fim switch Tokyo Tech Plac design.png|thumb|center|600px|Fig. 1. ''fim'' switch (Tokyo_Tech/B0010) design (Up:on state Down:off state)]]<br> | [[Image:Tokyo Tech fim switch Tokyo Tech Plac design.png|thumb|center|600px|Fig. 1. ''fim'' switch (Tokyo_Tech/B0010) design (Up:on state Down:off state)]]<br> | ||
Revision as of 08:35, 18 September 2015
fim switch[default ON](Tokyo_Tech/R0010)
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 the fim switch which has a J23119 promoter(BBa_K1632000) . Basically, the design of the fim switch(Tokyo_Tech) is similar to the fim switch(wild-type). The only difference is that we inserted restriction cut enzyme sites to fim switch(Tokyo_Tech). In detail, in fim switch(Tokyo_Tech) the sigma 70 promoter is exchanged to the J23119 promoter and there are two restriction enzyme cut sites inserted each in the front (SalI and BamHI) and in the back (BglII and MluI) of the promoter. Due to the insertion of the restriction enzyme cut sites, fim switch(Tokyo_Tech) has a promoter which can we exchanged with an arbitrary promoter. We actually changed J23119 promoter in the fim switch(Tokyo_Tech) to Lac promoter(BBa_B0010) as shown in the figure below.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 539
Illegal BamHI site found at 333 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]