BBa_K1361000
Based on the concept that change of cell motility by control of CheZ chemotaxis regulator could disturb pre-expressed curli fiber structure, we assume that if the inducer occurs, in this case acacia, CsgB protein will secrete into extracellular and act as nucleator to recruit soluble CsgA monomer into amyloid fibres, and the transcription of CheZ will be downgrade via cI-Plamda NOT gate. If induction condition disappears at a certain point, suppression of CheZ will be discharged. As a result, cells will be more moveable thus disturb or inhibit the establishment of curli structure. And in our final device, the dynamic detection could achieve by monitoring current change between two electrodes in the culture.
BBa_K1361001
This part contains the major subunit of curli fiber CsgA and its nucleator CsgB, in which CsgA is constitutive expression(a relatively weak promoter compared to BBa_K1361003) whereas CsgB is under the control of T7 promotor. This part cannot functioning alone without expression of CsgEFG genes in the same cell, for they coding the outer membrane channel secrete system for CsgA and CsgB.
BBa_K1361002
This part contains the major subunit of curli fiber CsgA and its nucleator CsgBtrunc, in which CsgA is constitutive expression whereas CsgBtrunc is under the control of Pbad promotor. CsgBtrunc is different from native CsgB for the deletion of C-terminal amino acid from 133 to 155 so that it cannot attach cell outer membrane. And CsgBtrunc itself is highly aggregative in the culture. This part is designed for immediate generation of large amount of curli fiber after adding the inducer. This part cannot function alone without the expression of CsgEFG genes in the same cell, for they coding the outer membrane channel secrete system for CsgA and CsgB.
BBa_K1361003
This part contains the major subunit of curli fiber CsgA and its nucleator CsgB, in which CsgA is constitutive expression(a relatively strong promoter compared to BBa_K1361001) whereas CsgB is under the control of T7 promoter. This part cannot function alone without expression of CsgEFG genes in the same cell, for they coding the outer membrane channel secrete system for CsgA and CsgB.
BBa_K1361004
This part contains the major subunit of curli fiber CsgA-his (CsgA modified by His tag) and its nucleator CsgB, in which CsgA-his is constitutive expression whereas CsgB is under the control of Pbad promoter. This part is designed for immediate generation of large amount of curli fiber after the adding of the inducer and the purification of CsgA by Ni-NTA reign or absorbing Au-NTA-Ni nano partial. This part cannot function alone without expression of CsgEFG genes in the same cell, for they coding the outer membrane channel secrete system for CsgA and CsgB.
BBa_K1361005
This part contains CsgE, CsgF, CsgG, which serve as the outer membrane secrete device for curli fiber, at relatively low constitutive. This part is for specific transport of CsgA(BBa_K1361999) and CsgB(BBa_K1361997) into extracellular matrix. CsgEFG(BBa_K1361992) was placed after a relatively weak constitutive promoter.
BBa_K1361006
This part contains the major subunit of curli fiber CsgA and its nucleator CsgBtrunc (a dissociative nucleator), in which CsgA is constitutive expression whereas CsgBtrunc is under the control of Pbad promoter. CsgBtrunc is different from native CsgB for the deletion of C-terminal amino acid from 133 to 155 so that it cannot attach cell outer membrane. And CsgBtrunc itself is highly aggregative in the culture. This part is designed for immediate generation of large amount of curli fiber after the adding of the inducer, and a stronger promoter for CsgA was chosen to parallel with BBa_K1361002. This part cannot function alone without expression of CsgEFG genes in the same cell, for they coding the outer membrane channel secrete system for CsgA and CsgB.
BBa_K1361007
This part contains the major subunit of curli fiber CsgA-his CsgA modified by His tag) and its nucleator CsgB, in which CsgA-his is constitutive expression whereas CsgB is under the control of Pbad promoter. This part is designed for immediate generation of large amount of curli fiber after adding of the inducer and the purification of CsgA by Ni-NTA reign, or absorbing Au-NTA-Ni nano partical. This part cannot function alone without expression of CsgEFG genes in the same cell, for they coding the outer membrane channel secrete system for CsgA and CsgB. |
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