Difference between revisions of "Part:BBa K395400:Experience"
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; Characterization of BBa_K395400
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<I>Tokyo Tech iGEM2010</I>
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In order to characterize K395400, LacI Mutant, we constructed BBa_K395401 combining I0500 and K082026(K395400) on pSB1A3 and used BBa_I20260 as a positive control and used promoterless gfp on pSB3K3 as a negative control.
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Furthermore, we constructed BBa_K395402 combining I0500 and BBa_I 732820 on pSB1A3 as a control experiment.
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As a GFP reporter, we used BBa_I7106 on pSB3K3.
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In order to measure the function of K395400), we introduced BBa_I7106 (lacI+pL-rbs-GFP-ter) on pSB3K3 and BBa_K395401/BBa_K395402 into DH5α. <br>
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<br>We confirmed that product of lacIM1 shows weaker repression to lac promoter than its wild type.<br>
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For more information, see [http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/lacIM1 our work in Tokyo_Tech 2010 wiki]
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[[Image:Tokyotech_LacIM1_data_ver2.png|center|thumb|300px|Figure4-2-2. Repression efficiency of LacIM1 (BBa_K395401) / LacIWT (BBa_K395402) exposed to arabinose and IPTG.  This work is done by Mitsuhiko Odera ]]
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Revision as of 20:36, 27 October 2010

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K395400

LacIM1 shows weaker repression than LacIWT because it has a lower affinity to lac promoter [1]. Though this part was registered by USTC(2008), it was not well characterized in their wiki. Therefore, we characterized this part.

We confirmed that lacIM1 shows weaker repression than LacIWT.(→[http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/lacIM1 more information])
We ligated BBa_K395400 together with Pbad/araC promoter. Then, we used it together with BBa_I7106(lacI+pL-RBS-GFP) on pSB3K3 (Fig. 3-2-1). Fluorescence was measured, and the result is shown in Fig. 3-2-2.


Figure3-2-1.
Figure3-2-2. Repression efficiency of LacIM1 (BBa_K395401) / LacIWT (BBa_K395402) exposed to arabinose and IPTG. This work is done by Mitsuhiko Odera


User Reviews

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Characterization of BBa_K395400

Tokyo Tech iGEM2010

In order to characterize K395400, LacI Mutant, we constructed BBa_K395401 combining I0500 and K082026(K395400) on pSB1A3 and used BBa_I20260 as a positive control and used promoterless gfp on pSB3K3 as a negative control. Furthermore, we constructed BBa_K395402 combining I0500 and BBa_I 732820 on pSB1A3 as a control experiment. As a GFP reporter, we used BBa_I7106 on pSB3K3. In order to measure the function of K395400), we introduced BBa_I7106 (lacI+pL-rbs-GFP-ter) on pSB3K3 and BBa_K395401/BBa_K395402 into DH5α.

We confirmed that product of lacIM1 shows weaker repression to lac promoter than its wild type.
For more information, see [http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/lacIM1 our work in Tokyo_Tech 2010 wiki]


Figure4-2-2. Repression efficiency of LacIM1 (BBa_K395401) / LacIWT (BBa_K395402) exposed to arabinose and IPTG. This work is done by Mitsuhiko Odera



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