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''' | ||
| + | [[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.
