Composite

Part:BBa_K1072003

Designed by: Yichong wang, Zhiwei li, Xihao liao and Junjie zhang   Group: iGEM13_SCUT   (2013-09-06)

Lux pL controlled LuxR+Lux pR autoinduceing luxI(lva tag)+Lux pR controlled Ndh

BBa_K1072003 is a composite device, including LuxI,LuxR and Ndh genes.

LuxR, controlled by Lux pI, generate LuxR protein, which will combined with AHL and activate Lux pR promotor. That complex increases the transcriptional rate of the lux pR promotor while also decrease the transceiptional rate of the Lux pL promoter.

LuxI is the generator of AHL controlled by Lux pR promoter.

Ndh was activated by AHL and LuxR complex and yields H2O2. When H2O2 enters the cell, it transiently changes its redox state and the native aerobic response control systems including ArcAB,which repress Lux pR normally, was inactived.

This part is an improvement of BBa_K546000, made by 2011 iGEM11_Wageningen_UR team. A Lux pR promoter, RBS, Ndh and double terminator has been added. This part can enhance the array quorum sensing.

We utilize the characteristic that H2O2 has absorbance at 240nm to complete the linear chart of standard H2O2 concentration as figure 1 by UV spectrophotometer.We choose part BBa_K546000 as the control for part BBa_K1072003 and get the concentration of H2O2 which is 7.21*10-2 mol/L.

Figure 1. the linear chart of standard H2O2 concentration


Figure 2. Transforming BBa_K1072003 into E.coli and expressing, we measure H2O2 OD under the control of BBa_K546000.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1821
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1101
    Illegal BsaI.rc site found at 1970


[edit]
Categories
//cds/enzyme
//direction/forward
Parameters
control(1) R0063 & (2) R0062 & (3) R0062
device_typefeedback
directionForward
latencySeconds
protein(1) LuxR (2) NADH dehydrogenase (3) luxI-LVA
signalling_molecule3OC6HSL (AHL)
switch_point(1) 2nM