Tokyo/Expression level check

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<br>[[Tokyo/Works|Works top]]  0.[[Tokyo/Works/Hybrid promoter|Hybrid promoter]]  1.[[Tokyo/Works/Formulation |Formulation]]  2.[[Tokyo/Works/Assay |Assay1]]  3.[[Tokyo/Works/Simulation |Simulation]]  4.[[Tokyo/Works/Assay2 |Assay2]]  5.[[Tokyo/Works/Future works |Future works]]
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'''assay2'''
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[[Tokyo/Activation check by cell-produced AHL |Activation check by cell-produced AHL ]]  [[Tokyo/Expression level check|Expression level check on ''promoters + plasmid sets'' of A and B sides]]
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==Expression level check on two different ''promoters + plasmid sets'' ==
==Expression level check on two different ''promoters + plasmid sets'' ==
==== Purpose: ====  
==== Purpose: ====  

Revision as of 06:34, 25 October 2007


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

assay2 Activation check by cell-produced AHL   Expression level check on promoters + plasmid sets of A and B sides

Expression level check on two different promoters + plasmid sets

Purpose:

To measure and compare the activities of two different promoters + plasmid sets by fluorescence of GFP downstream of each promoter. Lambda cI-regulated promoter and the lux lac hybrid promoter were tested.

Samples:

・A4Δp pc1-GFP
・A4 hybrid GFP PBR322TetR (+)AHL
・A4 hybrid GFP PBR322TetR (-)AHL
*The promoter pcI sequence was designed to contain OR1, -35, and -10 regions, but not R2 or OR3.

Procedure:

AHL assay Standard protocol
Wash
OD and fluorescence were measured 0, 3, and 6 hours after the fresh culture incubation started.

Result & Conclusion:

Two plasmid sets, A4Δp+pc1-GFP and A4 hybrid+GFP PBR322TetR (+)AHL, shows almost the same fluorescence of GFP, indicating that expression levels of both sets are almost the same though the latter is a bit smaller. Parameter set.jpg