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Volume 3,
Issue 1,
1968
Volume 3, Issue 1, 1968
- Articles
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Multiple Protein Functions in the Replication of Pox Virus DNA
More LessThe replication of pox virus DNA is known to require the prior synthesis of new proteins. It can thus be completely inhibited by the addition early after infection of either puromycin (Joklik & Becker, 1964) or p-fluorophenylalanine (FPA) (Appleyard & Zwartouw, 1965). This dependence upon protein synthesis has been related to a requirement for new enzymes and particularly the DNA polymerase which has been demonstrated in pox virus-infected cells (Magee, 1962; Green & Piña, 1962; Jungwirth & Joklik, 1965). Recently, however, Kates & McAuslan (1967) suggested that there is also need for a second type of protein function stoichiometrically related to DNA synthesis. We here draw attention to a difference in the action of puromycin and FPA which supports this interpretation and record additional findings which suggest that a third protein function may be needed to make ‘functional’ virus DNA.
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The Use of Density Gradient Centrifugation in a Zonal Centrifuge Rotor During the Purification of a Pear Virus
More LessStudies on the composition and structure of plant viruses can only be attempted after their purification from infected tissue homogenates. Owing to their intrinsic differences, however, the ease with which different viruses can be purified varies considerably. A few relatively stable viruses, e.g. tobacco mosaic virus or turnip yellow mosaic virus, can be treated with high concentrations of salt and precipitated by acid or alcohol without inactivation. With less stable viruses such methods are usually unsuccessful. The particles of such viruses, however, can be sedimented by ultracentrifugation and procedures developed frequently involve two treatments: (a) differential centrifugation and concentration after initial clarification of buffered homogenates such as with organic solvents (Steere, 1956; Tomlinson, Shepherd & Walker, 1959; Wetter, 1960), (b) further fractionation by rate or equilibrium density gradient centrifugation as developed by Brakke (1960). Because of the small capacity of the swinging-bucket rotor generally employed only relatively small volumes of the partially purified preparations can be fractionated in the gradients.
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