Difference between revisions of "Part:BBa K395401"

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<partinfo>BBa_K395401 short</partinfo>
 
<partinfo>BBa_K395401 short</partinfo>
  
In order to measure LacIM1(BBa_K395400), we designed this part.<br>
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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.
We confirmed that lacIM1 shows weaker repression than LacIWT.(→[http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/lacIM1 more information])<br>
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Furthermore, we constructed BBa_K395402 combining I0500 and BBa_I 732820 on pSB1A3 as a control experiment.
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.
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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 lacI proteins, 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 ]]
[[Image:Tokyotech_LacIM1_system_ver4.png|left|thumb|300px|Figure3-2-1. ]]
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[[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 ]]
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 19:33, 27 October 2010

pBad/araC-lacIM1

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 lacI proteins, 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

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961