Team:TUDelft/Module 3 How?

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='''How?'''=
='''How?'''=
From the literature review, two different genetic circuit configurations were contemplated: A [https://2009.igem.org/Team:TUDelft/Synthetic_Transcriptional_Cascade synthetic transcriptional cascade] approach, which has been showed to perform time-delay behavior in previous studies [[https://2009.igem.org/Team:TUDelft/References 4]] and an approach based on post-transcriptional regulation which we termed [https://2009.igem.org/Team:TUDelft/Biosynthetic_AND_gate biosynthetic AND gate].
From the literature review, two different genetic circuit configurations were contemplated: A [https://2009.igem.org/Team:TUDelft/Synthetic_Transcriptional_Cascade synthetic transcriptional cascade] approach, which has been showed to perform time-delay behavior in previous studies [[https://2009.igem.org/Team:TUDelft/References 4]] and an approach based on post-transcriptional regulation which we termed [https://2009.igem.org/Team:TUDelft/Biosynthetic_AND_gate biosynthetic AND gate].

Revision as of 20:51, 12 October 2009

How?

From the literature review, two different genetic circuit configurations were contemplated: A synthetic transcriptional cascade approach, which has been showed to perform time-delay behavior in previous studies [4] and an approach based on post-transcriptional regulation which we termed biosynthetic AND gate.

As the conjugation system will have two plasmids (conjugation and helper plasmids), the approach followed in this sub-project was to split the construction of the time-delay genetic circuit in two independent plasmids which in theory should be present in a single cell in order to initialize the time-delay. This can be achieved given two different selection markers and apply the two different selection pressures.

Due to the expected long delay time needed, an optimistic approach will be the combination of both approaches, synthetic transcriptional cascade and biosynthetic AND gate in order to gain a desire phenotype.

Figure 3. Time-delay genetic circuit approach. Two approaches have been considered: 1) (Blue circle) Synthetic transcriptional cascade, the delay is due to a sequential expression of genes, and 2) (Green circle) Biosynthetic AND gate, the delay is due to the need of the presence of two different events and post-transcriptional control. In order to achieve a long delay both approaches could be ultimately combined.


Which biobricks we could use

PartBiobrickWellPlatePlasmidAntibioticSize (bp)
pLac/PLacIR00101D1pSB1A2Amp200
PBlaI1401818N1pSB2K3Kan35
PlambclinI1200611J2pSB2K3Kan82
pTet/PtetR00406I1pSB1A2Amp54
key3cJ230083F1J23006Amp94
Lock3cJ230313L1J23006Amp42
GFPE004014K1pSB1A2Amp720
mRFP1E101018F1pSB2K3Kan681
cIC00514E1pSB1A2Amp750
RBSB00342M1pSB1A2Amp12
T (Double Terminator)B001523L1pSB1AK3Amp/Kan127
λp-RBS-GFP-TS0333585Box9pSB1A2Amp932
λp-RBS-mRFP1-TS0347379Box9pSB1A2Amp918
RBS - cI - RBSK08101312D2pSB1A2Amp819
TetRC00404A1pSB1A2Amp660

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