Two-dimensional display of lambda and Escherichia coli restriction fragments separated by Hg-Cs2SO4 centrifugation and gel electrophoresis |
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Authors: | H Yamagishi T Ikemura |
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Affiliation: | Department of Biophysics, Faculty of Science, Kyoto University, Kyoto 606 Japan |
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Abstract: | EcoRI restriction fragments derived from the DNA of bacteriophage lambda and Escherichia coli were fractionated by density gradient centrifugation of their mercury complexes in Cs2SO4 and subsequent electrophoresis on a horizontal agarose-gel slab. In this two-dimensional display, lambda fragments were resolved into six components and E coli fragments into more than 108 components. Bacterial chromosome regions contiguous to lambda prophage integrated at different sites were amplified by induction, and the EcoRI fragments were subjected to the two-dimensional analysis. As expected, the sets of amplified fragments were clearly different among the various lysogens. The approximate genome region affected by induction was estimated as one-tenth of the whole chromosome. |
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Keywords: | gene amplification by prophage induction G + C guanine + cytosine kb kilobase pairs TCG Tris-Casamino acid glucose |
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