Difference between revisions of "Part:BBa K1632023"
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<partinfo>BBa_K1632023 short</partinfo>
 
<partinfo>BBa_K1632023 short</partinfo>
  
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At the first stage of wet experiment, we used the cells which have the plasmid which is Pcon_rhlR_TT_Plux_rbs_cmR to characterize the function of rbs_cmR(<partinfo>BBa_K395160</partinfo> by iGEM10_Tokyo_Tech). But the cells showed active growth even in the absence of AHL (Fig. 1.The cells growth with Cm)
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At the first stage of wet experiment, we used the cells which have the plasmid which is Pcon_rhlR_TT_Plux_rbs_CmR to characterize the function of rbs_CmR(<partinfo>BBa_K395160</partinfo> by iGEM10_Tokyo_Tech). But the cells showed active growth even in the absence of AHL (Fig. 1.The cells growth with Cm)
 
   
 
   
 
[[Image:Tokyo_Tech Pcon_rbs_rhlR_TT_Plux_rbs_cmR.png|thumb|center|550px|<b>Fig. 1.</b> The cells growth with Cm]]
 
[[Image:Tokyo_Tech Pcon_rbs_rhlR_TT_Plux_rbs_cmR.png|thumb|center|550px|<b>Fig. 1.</b> The cells growth with Cm]]
  
 
From the results of this experiment, initially designed circuits showed leaky expression of CmR.  So we inserted an ssrA degradation tag to the C-terminal of CmR.<br>
 
From the results of this experiment, initially designed circuits showed leaky expression of CmR.  So we inserted an ssrA degradation tag to the C-terminal of CmR.<br>
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In the experiment using the Pcon_lasR_TT_Plux_cmRssrA (BBa_K1632022) and Pcon_rhlR_TT_Plux_cmRssrA (BBa_K1632023), we could not observe cell growth for cells that owned the ssrA-taged plasmid, in the absence of AHL<br>
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In the experiment using the Pcon_lasR_TT_Plux_CmRssrA (BBa_K1632022) and Pcon_rhlR_TT_Plux_CmRssrA (BBa_K1632023), we could not observe cell growth for cells that owned the ssrA-taged plasmid, in the absence of AHL<br>
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From our experiment, cmRssrA is confirmed work better than CmR without ssrA tag for our project.
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From our experiment, CmRssrA is confirmed work better than CmR without ssrA tag for our project.
  
 
For more information, see [http://2015.igem.org/Team:Tokyo_Tech/Project our work in Tokyo_Tech 2015 wiki].
 
For more information, see [http://2015.igem.org/Team:Tokyo_Tech/Project our work in Tokyo_Tech 2015 wiki].

Revision as of 00:16, 18 September 2015

J23100_rbs_rhlR_TT_Plux_rbs_CmRssrA

At the first stage of wet experiment, we used the cells which have the plasmid which is Pcon_rhlR_TT_Plux_rbs_CmR to characterize the function of rbs_CmR(BBa_K395160 by iGEM10_Tokyo_Tech). But the cells showed active growth even in the absence of AHL (Fig. 1.The cells growth with Cm)

Fig. 1. The cells growth with Cm

From the results of this experiment, initially designed circuits showed leaky expression of CmR. So we inserted an ssrA degradation tag to the C-terminal of CmR.
In the experiment using the Pcon_lasR_TT_Plux_CmRssrA (BBa_K1632022) and Pcon_rhlR_TT_Plux_CmRssrA (BBa_K1632023), we could not observe cell growth for cells that owned the ssrA-taged plasmid, in the absence of AHL
From our experiment, CmRssrA is confirmed work better than CmR without ssrA tag for our project.

For more information, see [http://2015.igem.org/Team:Tokyo_Tech/Project our work in Tokyo_Tech 2015 wiki].

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 301
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 776
    Illegal BsaI.rc site found at 933