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14 June 2004 Making temporal maps using bacterial luciferase: Bacteriophage
Jonathan Kuhn, Rachel Broza, Ekaterina Verkin
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Proceedings Volume 5329, Genetically Engineered and Optical Probes for Biomedical Applications II; (2004) https://doi.org/10.1117/12.530706
Event: Biomedical Optics 2004, 2004, San Jose, CA, United States
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.
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Jonathan Kuhn, Rachel Broza, and Ekaterina Verkin "Making temporal maps using bacterial luciferase: Bacteriophage", Proc. SPIE 5329, Genetically Engineered and Optical Probes for Biomedical Applications II, (14 June 2004); https://doi.org/10.1117/12.530706
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KEYWORDS
Proteins
Resistance
Organisms
Bacteria
Pathogens
Genetics
Polymers
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