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

 
(→‎User Reviews)
Line 19: Line 19:
 
<!-- End of the user review template -->
 
<!-- End of the user review template -->
 
<!-- DON'T DELETE --><partinfo>BBa_K143010 EndReviews</partinfo>
 
<!-- DON'T DELETE --><partinfo>BBa_K143010 EndReviews</partinfo>
 +
 +
 +
{|width='80%' style='border:1px solid gray'
 +
|-
 +
|width='10%'|
 +
<partinfo>BBa_K143010 AddReview 0</partinfo>
 +
<I>Username</I>
 +
|width='60%' valign='top'|
 +
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.
 +
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.
 +
https://static.igem.org/mediawiki/parts/4/4e/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.
 +
|};

Revision as of 01:42, 27 September 2013


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K143010

User Reviews

UNIQ2ef6bcc6a47e0ff1-partinfo-00000000-QINU UNIQ2ef6bcc6a47e0ff1-partinfo-00000001-QINU


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.

;