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Characterization of the Bacillus stearothermophilus phage φμ-4 and Its DNA

Virology, ISSN: 0042-6822, Vol: 41, Issue: 1, Page: 91-100
1970
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Bacteriophage φμ-4 was purified from lysates by isoelectric precipitation and CsCl gradient centrifugation. This phage displays an Escherichia coli phage lambda-like appearance in electron microscopy with polyhedrical heads and long tails, but without terminal plates and tail fibers. Bacteriophage φμ-4 is a DNA virus. The DNA has a molecular weight 26 × 10 6 daltons, a buoyant density of 1.719 g/cm 3, a G + C-content of 58% and appears to contain no unusual base residues. The DNA strands can be separated by alkaline CsCl gradient centrifugation or annealing to poly U,G, and subsequent CsCl gradient centrifugation. DNA of phage φμ-4 is an excellent template for in vitro transcription by both E. coli RNA-polymerase and host RNA-polymerase ( Bacillus stearothermophilus ). There are 15 polymerase binding sites on μμ-4 DNA for both enzymes as determined bv the heparin technique. φμ-4 DNA is transcribed completely in vitro. In vitro synthesized RNA is, however, not as well translated as other templates in a cell-free E. coli system.

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