Difference between revisions of "Part:BBa K1632020"

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<partinfo>BBa_K1632020 short</partinfo>
 
<partinfo>BBa_K1632020 short</partinfo>
  
<span style="margin-left: 10px;">In our project, we wanted to use the ''CmR''. 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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<span style="margin-left: 10px;">In our project, we wanted to use the ''CmR''. At the first stage of wet experiment, we used the cells which have the plasmid which is Pcon_''rhlR''_TT_Plux_rbs_''CmR''(Fig. 1. Our initially design of a part containing a previously existing part) 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. 2.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]]
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[[Image:Tokyo_Tech Pcon_rhlR_TT_Plux_cmR.png|thumb|center|600px|<b>Fig. 1.</b>Our initially design of a part containing a previously existing part]]
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[[Image:Tokyo_Tech Pcon_rbs_rhlR_TT_Plux_rbs_cmR.png|thumb|center|550px|<b>Fig. 2.</b> The cells growth with Cm]]
  
 
<span style="margin-left: 10px;">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>
 
<span style="margin-left: 10px;">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>

Revision as of 18:55, 18 September 2015

rbs_CmR(ssrA degradation tag)

In our project, we wanted to use the CmR. At the first stage of wet experiment, we used the cells which have the plasmid which is Pcon_rhlR_TT_Plux_rbs_CmR(Fig. 1. Our initially design of a part containing a previously existing part) 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. 2.The cells growth with Cm)

Fig. 1.Our initially design of a part containing a previously existing part
Fig. 2. 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.

Fig. 2. The cells which have BBa_K1632022 growth with Cm
Fig. 3. The cells which have BBa_K1632023 growth with Cm


In the experiment using the Pcon_lasR_TT_Plux_CmRssrA (BBa_K1632022) (Fig.2 The cells which have BBa_K1632022 growth with Cm) and Pcon_rhlR_TT_Plux_CmRssrA (BBa_K1632023) (Fig.3 The cells which have BBa_K1632023 growth with Cm), 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
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]