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Proceedings Paper

Making temporal maps using bacterial luciferase: Bacteriophage
Author(s): Jonathan Kuhn; Rachel Broza; Ekaterina Verkin
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Paper Abstract

A method for making temporal maps in bacteria, plasmids and bacteriophages is described. A cassette containing both the genes for bacterial luciferase and kanamycin resistance can be introduced at precise sites. The technique involves clonging followed by genetic recombination. The result is formation of structures that have the luciferase genes in place of the normal DNA and this allows the very precise measurement of transcription/translation of the substituted regions. Very low levels of transcription as well as the kinetics of induction can be easily ascertained. As a specific demonstration of this general method, the technique was used with bacteriophage λ, one of the best known organisms. By measuring light emission, the expression of luciferase was followed after induction for both early and late genes. The exact timing of initial expression of genes was also determined by sampling at very short intervals. The results show that the early genes express almost without delay implying that the function of the N antitermination system is not temporal regulation.

Paper Details

Date Published: 14 June 2004
PDF: 12 pages
Proc. SPIE 5329, Genetically Engineered and Optical Probes for Biomedical Applications II, (14 June 2004); doi: 10.1117/12.530706
Show Author Affiliations
Jonathan Kuhn, Technicon-Israel Institute of Technology (Israel)
Rachel Broza, Technicon-Israel Institute of Technology (Israel)
Ekaterina Verkin, Technicon-Israel Institute of Technology (Israel)

Published in SPIE Proceedings Vol. 5329:
Genetically Engineered and Optical Probes for Biomedical Applications II
Alexander P. Savitsky; Darryl J. Bornhop; Ramesh Raghavachari; Samuel I. Achilefu, Editor(s)

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