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Help:Terminators/Measurement
From partsregistry.org
Each termination efficiency value indicated in the terminator part tables is followed by a two or three letter initials in brackets of the individual who made that measurement. The terminator efficiency characterization work done to date is summarized here so that you can reuse and improve upon previous efforts to characterize terminator function. Jason, Cassie and Caitlin all took similar approaches to measuring termination efficiency. They used a bicistronic, dual fluorescent protein to measure termination efficiency. By comparing the fluorescence ratio of the two reporters in the absence and presence of a terminator in between, you can assess the termination efficiency. In the absence of a terminator, the fluorescence ratio is normalized to 1. A perfect terminator with 100% termination efficiency should result in no fluorescence from the second reporter.
Click the links below to learn more about Jason's, Cassie's, Caitlin's or Chris' approach to characterizing termination efficiencies.
- [JK] - Measurements done by Jason Kelly in Drew Endy's lab
- [CH] - Measurements done by Cassie Huang in Tom Knight's lab
- [CC] - Measurements done by Caitlin Conboy in Drew Endy's lab
- [JCA] - Measurement done by Chris Anderson
References
- 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 pmid:12080425.
- 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 pmid:12402322.
- Choe J, Guo HH, and van den Engh G. A dual-fluorescence reporter system for high-throughput clone characterization and selection by cell sorting. Nucleic Acids Res 2005 Mar 14; 33(5) e49. doi:10.1093/nar/gni049 pmid:15767274.
- Nicholson AW. Function, mechanism and regulation of bacterial ribonucleases. FEMS Microbiol Rev 1999 Jun; 23(3) 371-90. pmid:10371039.
- Nojima T, Lin AC, Yamamoto T, Endo I, and Fujii T. Controlling the expression ratio of two proteins by inserting a terminator between the two genes. Nucleic Acids Symp Ser (Oxf) 2006 329-30. doi:10.1093/nass/nrl164 pmid:17150951.
