Difference between revisions of "Part:BBa K395400:Experience"

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Applications of BBa_K395400

User Reviews

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UNIQd5a28a117e826d48-partinfo-00000002-QINU

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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 ===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.<br><br>
-We confirmed that lacIM1 shows weaker repression than LacIWT.(→[http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/lacIM1 more information])<br>
-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.
-[[Image:Tokyotech_LacIM1_system_ver4.png|left|thumb|300px|Figure3-2-1. ]]
-[[Image:Tokyotech_LacIM1_data_ver2.png|center|thumb|300px|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===