Composite

Part:BBa_K557002

Designed by: Yinghong Lan   Group: iGEM11_USTC-China   (2011-10-02)
Revision as of 21:31, 5 October 2011 by Pythonwawa (Talk | contribs) (→‎Experimental Data)

Toggle-switch+Aptamer+CheZ

This part is the assembly of riboswitches followed by an important gene CheZ in keeping E.coli motile (BBa_K557001) finely tuned by small molecules and toggle switches (BBa_K228003) pushed on and off while memorizing current state. It would make it possible that bacterial colonies will automatically divide into two parts after certain time, during which riboswitches would act as the main power to drive two parts away from each other while toggle switches would play the role of giving birth to two 'different' kinds of bacteria in one colony.
With the help of BBa_K557003, it can produce more balanced colony division.
NO1.png
NO2.png
NO3.jpg

Model

Toggle-switch1.jpg
Figure 1: Constructions to modulate the ratio of toggle switches. The above bar represents pSB3T5, a low to medium copy BioBrick standard vector, where LuxpR (BBa_R0062) controls the expression of cI repressor which would influence the ratio of toggle switch. (Images by USTC-IGEM with TinkerCell)

Experimental Data

Test of Incorporated Aptamer-cheZ part into one side of the original Toggle-switch

650px-Resul.jpg

Figure5.The growing state of the reprogrammed bacteria with original Toggle-switch-Aptamer-cheZ Device(Red circle:the range of motion, Blue Triangle:the gradient of thepphylline)

300px-Resddul.jpg

Figure6.The fluorescent Photo(4X Objective) of the colony shown in Figure5 Left.

    According to the results above, by using this device we can not make the reprogrammed bacteria be divided into two different states effectively, and because of the bias to the green fluorescence and moving towards the high concentration of the theophylline, the bacteria trending to stay are very rare.

Acknowledgements

Here we want really want to thank the Professor J. S. Parkinson for providing ΔcheZ strain RP1616 and the corresponding wild type RP437 and Professor J. P. Gallivan for directing our construction of riboswitches.

Reference

1.Shana Topp, Justin P. Gallivan (2006) Guiding Bacteria with Small Molecules and RNA. J.Am.Chem.Soc, 129,6870-6811

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 10
    Illegal AgeI site found at 122
  • 1000
    COMPATIBLE WITH RFC[1000]

[edit]
Categories
Parameters
functionallow bacterial colonies to automatically divide into two parts after certain time