Difference between revisions of "Part:BBa K1412001"
(→‎protocol)
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== '''protocol''' ==
 
== '''protocol''' ==
 
1.Transformation
 
1.Transformation
 +
 
2.Extract plasmids
 
2.Extract plasmids
 +
 
3.Digestion
 
3.Digestion
 +
 
4.DNA gel electrophoresis
 
4.DNA gel electrophoresis
 +
 
5.Gel Extraction
 
5.Gel Extraction
 +
 
6.ligation
 
6.ligation
 +
 
7.Transformation
 
7.Transformation
 +
 
8.Extract plasmids
 
8.Extract plasmids
 +
 
9.Digestion
 
9.Digestion
 +
 
10.DNA gel electrophoresis
 
10.DNA gel electrophoresis
 +
 
11.Transform the plasmid into CL-1
 
11.Transform the plasmid into CL-1
 +
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 09:52, 5 September 2014

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


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


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


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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*]