Purification of Bacteriophages and SDS-PAGE Analysis of Phage Structural Proteins from Ghost Particles
作者： Pascale Boulanger
DOI: 10.1007/978-1-60327-565-1_13
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摘要: Concentration and purification of infectious particles are prerequisites for structural functional characterization bacteriophages. The methods detailed in the first part this chapter outline protocols commonly used to obtain purified phages: concentration phage by precipitation with polyethylene glycol their centrifugation CsCl step gradients subsequently equilibrium centrifugation. This sequence procedures, if carried out as a whole, ensures high quality, which is well suited most analytical techniques characterize bacteriophage particles. second describes preparation "ghosts" or DNA-less These should be preferred entire bacteriophages one-dimensional SDS-PAGE analysis proteins, since running proteins through gel not disturbed presence DNA. allows an optimal resolution, necessary proteomic approaches such N-terminal protein sequencing mass spectrometry using isolated from distinct bands.
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