Revision as of 14:29, 4 October 2014

Basic part of biobrick BBa_K1412001

What it is

Compared to part BBa_1412001, this part substitutes BBa_R0040, a constant expression promoter in our engineered bacteria CL-1, for BBa_K206000.

What it does

As a control group for K1412001, we use this part to eliminate background expression of AraC, and it’s also capable to form ellipse on the plate under the induction of IPTG.

How to use it

Transforming this part into CL-1, we got the engineering bacteria. Incubate them on the plate with fit concentration of IPTG and chloromycetin, we observed the chemotaxis diameter over time, and got the experimental data. It is also used to form ellipse.

Experimental Data

With K20600(pBAD) as an inhibit promoter

By adding chloramphenicol of different concentrations. Team XMU2014 found the best chemotaxis as the concentration is 50μg/ml

Team XMU2014 set the concentration of chloramphenicol at 50ug/ml, by adding IPTG of different concentration levels, Team XMU2014 found that when the concentration of IPTG lies between 0.02 and 0.075, the chemotaxis diameter is better than the others.

Team XMU2014 set the concentration of chloramphenicol at 50μg/ml and the concentration of IPTG at 0.025mM, and found the minimum inhibition concentration of Ara is 0.2% and best inducing concentration is 0.02%.

Team XMU2014 set the concentration of IPTG and Ara on the plate at 0.025mM and 0.02% deliberately, then choose two points 0.5cm (up) [1cm (down)] away form each other on the plate. Team XMU2014 achieved the goal of command the CL-1 to form hyperbolic curve by doting 0.25mM IPTG（√） and 1% Ara （X）on the two points deliberately.

protocol

1.Transformation

2.Extract plasmids

3.Digestion

4.DNA gel electrophoresis

5.Gel Extraction

6.Ligation

7.Transformation

8.Extract plasmids

9.Digestion

10.DNA gel electrophoresis

11.Transform the plasmid into CL-1

Sequence and Features