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Part:BBa_K1412001

Designed by: Tangduo Zhang   Group: iGEM14_XMU-China   (2014-08-28)
Revision as of 14:02, 1 October 2014 by Tam.Zhang (Talk | contribs) (→‎Experimental data)

Endow the E.coli engineered strain(CL-1) the ability of chemotaxis


What it is

Composite part enables the chemotaxis of our engineering bacteria could be regulated by IPTG and L-Arabinose; pBAD and pLac promoters control the expression of Lac I and CheZ.


What it does

when L-Arabinose is added, pBAD promoter is activated, expressing Lac I, inhibiting the expression of CheZ. When IPTG is added, IPTG combines with Lac I, forming a complex, thus relief the inhibition and the CheZ express, make the ability of chemotaxis recover.


How to use it in their project

Use IPTG as an inducer and L-Arabinose as an inhibitor to take control of the chemotaxis of the engineering bacteria.


Experimental data

More information in linking page of the following pictures.

Curve of chemotaxis diameter under different antibiotics concentrations.png


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

Curve of chemotaxis diameter over time under different IPTG concentration .png


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.


Curve of chemotaxis diameter over time under different Ara concentration .png


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%.


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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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Reference

[1] [http://www.ncbi.nlm.nih.gov/pubmed/21998392/ Sequential Establishment of Stripe Patterns in an Expanding Cell Population Chenli Liu et al.Science 334, 238 (2011);DOI: 10.1126/science.1209042]

[2][http://www.biomedsearch.com/nih/Reprogramming-bacteria-to-seek-destroy/20453864.html Reprogramming bacteria to seek and destroy an herbicide Joy Sinha1, Samuel J Reyes1 & Justin P Gallivan1*]


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