Difference between revisions of "Part:BBa K1949101"

(Ⅰ.Adjustment of MazF Expression)
Line 22: Line 22:
 
=====result=====
 
=====result=====
 
<span style="margin-left: 10px;">It was found that the cell growth is inhibited when the concentration of arabinose, the inducer of MazF, is more than 0.02%. However, when arabinose concentration is 0.2%, GFP fluorescence intensity falls markedly.
 
<span style="margin-left: 10px;">It was found that the cell growth is inhibited when the concentration of arabinose, the inducer of MazF, is more than 0.02%. However, when arabinose concentration is 0.2%, GFP fluorescence intensity falls markedly.
 +
 +
[[Image:Arabinose.png|thumb|center|600px|Fig. 1. Relative value of Turbidity, RFU of GFP and RFU of GFP / Turbidity of medium where E. coli was cultured vs concentration of arabinose.]]<br>
  
 
====Ⅱ.<i>mazEF</i> System Assay ~Stop & GO~====
 
====Ⅱ.<i>mazEF</i> System Assay ~Stop & GO~====

Revision as of 08:19, 17 October 2016


PBAD-rbs-mazF

Characterize

We performed experiment to work final genetic circuits.

This experiment consists of the four parts below.

Ⅰ.Adjustment of MazF Expression

Ⅱ.mazEF System Assay ~Stop & GO~

Ⅲ.mazEF System Assay ~Go & Stop~

Ⅳ.Control of Cell Growth

Ⅰ.Adjustment of MazF Expression

In order to control the TA system well, it is necessary to adjust the expression of MazF as a toxin. It has been reported that MazF has very strong ability to inhibit cell growth, and antitoxins cannot recover it under overexpression of MazF. Therefore, we explored the conditions that can control the TA system by adjusting the expression of MazF(Fig. 1.).


result

It was found that the cell growth is inhibited when the concentration of arabinose, the inducer of MazF, is more than 0.02%. However, when arabinose concentration is 0.2%, GFP fluorescence intensity falls markedly.

Fig. 1. Relative value of Turbidity, RFU of GFP and RFU of GFP / Turbidity of medium where E. coli was cultured vs concentration of arabinose.

Ⅱ.mazEF System Assay ~Stop & GO~

result

Ⅲ.mazEF System Assay ~Go & Stop~

result

Ⅳ.Control of Cell Growth

result

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