Endy:Screening plasmid

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This project is in progress -- results described are preliminary. If you would like to keep up to date with this work, please subscribe to the RSS feed.

Introduction

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.


Current Experimental Work

Screening Plasmid 0.X

Screening Plasmid 1.0

This is the current working version of the screening plasmid. It is available from the MIT Registry of Standard Biological Parts as pSB1A10.

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:

Documents

References

  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