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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.
Screening Plasmid 1.0
Characterization of Empty Screening Plasmid 1.0
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).
Terminator Characterization via Screening Plasmid 1.0
Inverter Characterization via 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:
- /Registry technical report/ (in progress)
- 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 |
- 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 |