Difference between revisions of "Part:BBa K143010:Experience"

 
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We put amilCP(<bbpart>BBa_K592009</bbpart>) as a reporter under the promoter ctc to expect a visual signal when B''.subtilis'' grow to the post-log phase. But In our experiment, we failed to see obviously blue during all of the grow phases. We have tried almost all of the methods recommended by previous team(<bbpart>BBa_K143037</bbpart>)(42℃ culture temperature, additional of NaCl,ethanol and blue light ) to trigger the physical stress response of engineered bacterial, but experiment group still show no differences with the negative control.
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To explore the possible reason of these results, Many work of literature research have been done. It seems that the expressing level of promoter ctc is somewhat low as to our requirement. So we think the  promoter ctc may be too weak to express amilCP for visible blue.
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In view of this, we have come up a idea to  design a positive feedback system like below.
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https://static.igem.org/mediawiki/parts/4/4e/USTC_China_iGEM13_BBa_K1074007_1.JPG
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In this circuit, we put sigma B factor under the promoter ctc with amilcp. When the colony enter the post-log phase, the promotor ctc is activated, then more sigma B factor is produced, further activites promoter ctc.  We hope this positive feedback could increase the expression of Promoter ctc to get a visible signal of amilCP.
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Revision as of 01:42, 27 September 2013


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Username

We put amilCP(BBa_K592009) as a reporter under the promoter ctc to expect a visual signal when B.subtilis grow to the post-log phase. But In our experiment, we failed to see obviously blue during all of the grow phases. We have tried almost all of the methods recommended by previous team(BBa_K143037)(42℃ culture temperature, additional of NaCl,ethanol and blue light ) to trigger the physical stress response of engineered bacterial, but experiment group still show no differences with the negative control. To explore the possible reason of these results, Many work of literature research have been done. It seems that the expressing level of promoter ctc is somewhat low as to our requirement. So we think the promoter ctc may be too weak to express amilCP for visible blue. In view of this, we have come up a idea to design a positive feedback system like below. USTC_China_iGEM13_BBa_K1074007_1.JPG

In this circuit, we put sigma B factor under the promoter ctc with amilcp. When the colony enter the post-log phase, the promotor ctc is activated, then more sigma B factor is produced, further activites promoter ctc. We hope this positive feedback could increase the expression of Promoter ctc to get a visible signal of amilCP.

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