Endy:Screening plasmid/v1.0: Difference between revisions
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==Design== | |||
[[Image:ScreeningPlasmid1.0.PNG|600px|left|thumb|'''Design of Screening Plasmid 1.0:''' We are using the Pbad arabinose-inducible induction system <cite>Khlebnikov</cite> as a tunable input. GFP is a measure of input and RFP is a measure of output. A Biobricks cloning site enables easy insertion of any Biobricks part. RNase E sites create independence between the mRNA stability of the device being screened and the mRNA stability of the fluorescent proteins. In particular, we suspect mRFP1 contains internal RNaseE cut sites and have added a hairpin 5’ of the coding region to slow degradation by RNase E. <cite>Smolke</cite>]] | [[Image:ScreeningPlasmid1.0.PNG|600px|left|thumb|'''Design of Screening Plasmid 1.0:''' We are using the Pbad arabinose-inducible induction system <cite>Khlebnikov</cite> as a tunable input. GFP is a measure of input and RFP is a measure of output. A Biobricks cloning site enables easy insertion of any Biobricks part. RNase E sites create independence between the mRNA stability of the device being screened and the mRNA stability of the fluorescent proteins. In particular, we suspect mRFP1 contains internal RNaseE cut sites and have added a hairpin 5’ of the coding region to slow degradation by RNase E. <cite>Smolke</cite>]] | ||
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Revision as of 21:55, 23 April 2007
Design
Characterization of Empty Screening Plasmid 1.0
- Endy:Screening plasmid/v1.0/Hairpin - characterize effectiveness of the hairpin to stabilize the mRFP1 transcript
- Endy:Screening plasmid/v1.0/RNAseE sites - characterize effectiveness of the RNAseE sites
To ensure our screening system was operating as expected we measured the expression levels of GFP and mRFP1 when the plasmid did not contain a part (e.g. input equals output).
Measurement of Parts and Devices using Screening Plasmid 1.0
Terminator Characterization via Screening Plasmid 1.0
Inverter Characterization via Screening Plasmid 1.0
Inverter library screening via Screening Plasmid 1.0
To be added.