Difference between revisions of "Part:BBa K1949101"

Revision as of 08:27, 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.

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.).

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~

The biggest attraction of the TA system is that it is able to control cell growth and synthesis of protein. In this experiment, MazE expression was induced by the addition of IPTG(2 mM) after MazF expression was induced by the addition of arabinose(0.02%). As a result, it was able to resuscitate from a state of being inhibited cell growth. We named this experiment as "Stop & Go" because it was to resuscitate growth from inhibiting cell growth.

result

Ⅲ.mazEF System Assay ~Go & Stop~

result

Ⅳ.Control of Cell Growth

result

Sequence and Features