Difference between revisions of "Endy:Screening plasmid"

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(Screening Plasmid 1.0)
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==Screening Plasmid 1.0==
==Screening Plasmid 1.0==
[[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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*[[Endy:Screening plasmid 1.0/Background|Background]]
*[[Endy:Screening plasmid 1.0/Protocols|Protocols]]
*[[Endy:Screening plasmid status|Status]]
*[[Endy:Screening plasmid constructs|Constructs]]
===Characterization of Empty Screening Plasmid 1.0===
*[[Endy:Screening plasmid/v1.0]]
*[[Endy:Screening plasmid/v1.0]]

Revision as of 21:52, 23 April 2007

This page is a work in progress.


Construction of engineered biological systems from collections of standard biological parts requires mechanisms for rapid and reliable characterization of parts. Additionally, the difficulty of rational part design necessitates library screening systems that can be employed in service of tuning part performance. Here we describe pSB1A10, a system for both characterizing and screening transcription-based parts based on their input / output function. We demonstrate the successful operation of this system by characterizing and tuning genetic inverters and transcriptional terminators.

Schematic of the screening plasmid design. It consists of 4 components: (1) Tunable input (2) Input measurement (3) Part/Device insertion site (4) Output measurement.

Screening Plasmid 1.0

Screening Plasmid 1.5

This version of the screening makes use of an AHL-based induction system (<bbpart>F2620</bbpart>).

Screening Plasmid 2.0

We are in the process of designing a new version of the screening plasmid to account for some of the shortcomings of the previous version. Details can be found here:



  1. Khlebnikov A, Skaug T, and Keasling JD. Modulation of gene expression from the arabinose-inducible araBAD promoter. J Ind Microbiol Biotechnol. 2002 Jul;29(1):34-7. DOI:10.1038/sj.jim.7000259 | PubMed ID:12080425 | HubMed [Khlebnikov]
  2. Smolke CD and Keasling JD. Effect of gene location, mRNA secondary structures, and RNase sites on expression of two genes in an engineered operon. Biotechnol Bioeng. 2002 Dec 30;80(7):762-76. DOI:10.1002/bit.10434 | PubMed ID:12402322 | HubMed [Smolke]

All Medline abstracts: PubMed | HubMed