Tokyo/Expression level check
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<br>・A4 hybrid GFP PBR322TetR (+)AHL | <br>・A4 hybrid GFP PBR322TetR (+)AHL | ||
<br>・A4 hybrid GFP PBR322TetR (-)AHL | <br>・A4 hybrid GFP PBR322TetR (-)AHL | ||
+ | <br>*The promoter pcI sequence was designed to contain OR1, -35, and -10 regions, but not R2 or OR3. | ||
==== Procedure: ==== | ==== Procedure: ==== |
Revision as of 03:40, 25 October 2007
Contents |
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.