Tokyo/Works/Assay2

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==[[Tokyo/Expression level check|Expression level check on ''promoters + plasmid sets'' of A and B sides]]==
==[[Tokyo/Expression level check|Expression level check on ''promoters + plasmid sets'' of A and B sides]]==
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<br>Two plasmid sets, A4 ΔP/pc1-GFP(upper side of Fig.2, off/on respectively) and A4 hybrid-promoter(+AHL to activate)/pBR322tetR (lower side of Fig.2, on/off respectively), showed almost the same fluorescence of GFP. This result indicates that expression levels of both sets were almost the same, though the latter was a bit smaller.[[Image:pC1GFP.jpg|thumb|300px| '''Fig.3''' Genes downstream of the promoters on the both sides were expressed to almost the same degrees in terms of GFP fluorescence.]]
<br>Two plasmid sets, A4 ΔP/pc1-GFP(upper side of Fig.2, off/on respectively) and A4 hybrid-promoter(+AHL to activate)/pBR322tetR (lower side of Fig.2, on/off respectively), showed almost the same fluorescence of GFP. This result indicates that expression levels of both sets were almost the same, though the latter was a bit smaller.[[Image:pC1GFP.jpg|thumb|300px| '''Fig.3''' Genes downstream of the promoters on the both sides were expressed to almost the same degrees in terms of GFP fluorescence.]]
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==== ==> see more [[Tokyo/Expression level check|details]]====
==== ==> see more [[Tokyo/Expression level check|details]]====

Revision as of 03:13, 27 October 2007


Works top   0.Hybrid promoter   1.Formulation   2.Assay1   3.Simulation   4.Assay2   5.Future works

Activation check by cell-produced AHL    Expression level check on promoters + plasmid sets of A and B sides

Objective

Parameters for the equations in Formulation have been experimentally determined in Assay1. Analysing the result of the simulations, the following experiments were turned out to be necessary.

Activation check by cell-produced AHL

Fig.1 By culturing with AHL producing cells, whether the lux lac hybrid promoter was activated without externally adding AHL. Regardless of the copy numbers of the luxI encoding plasmids, the hybrid promoter was activated.

Objective

To check whether worker E. coli (Sender) can produce enough amount of AHL for our model to work by using different copy numbers of plasmids


Result & Conclusion

Not only high copy number plasmid pSB1, but also low copy number plasmid pSB4 and pBR produced enough AHL to activate the LacI hybrid promoter in other cells. Especially, pBR remarkably produced AHL in the present experiment.


==> see more details







==> see more details