Difference between revisions of "Part:BBa K082005:Experience"

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<I>Tokyo Tech iGEM2010</I>
 
<I>Tokyo Tech iGEM2010</I>
 
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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.
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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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We confirmed that lacIM1 shows weaker repression than LacIWT.
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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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[[Image:Tokyotech_LacIM1_system.png|left|thumb|200px|Figure3-2-1. ]]
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As a GFP reporter, we used BBa_I7106 on pSB3K3.
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[[Image:Tokyotech_LacIM1_data.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 ]]
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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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(→[http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/lacIM1 more information])
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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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Latest revision as of 20:32, 27 October 2010

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Applications of BBa_K082005

User Reviews

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Characterization of BBa_K082005

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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