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Closed Access item The orderly, in vitro emergence of DNA from bacteriophage φ29 particles

Authors:Krawiec, S.
Jiménez, Fernando
García, Juan Antonio
Villanueva, Nieves
Sogo, José M.
Salas, Margarita
Issue Date:2-Jun-1981
Publisher:Elsevier
Citation:Virology 111(2): 440-454 (1981)
Abstract:φ29 DNA-containing 12−13− particles (produced by infecting nonsuppressor hosts of Bacillus subtilis with phage containing suppressible mutations in cistrons 12 and 13) can be complemented with lysates containing proteins p12* and p13 to yield infectious phage. Complementation of these particles with lysates containing p12* but not p13 or complementation with purified p12* in the absence of p13 produces a structure (called complex) which has a markedly different organization. Electron microscopy and sedimentation analysis after digestion with DNase I or proteinase K indicate that complex is composed of an intact phage head with a genome-sized linear DNA molecule attached at the collar-tail region. EcoRI digestion establishes that the DNA molecule has a unique orientation. Gel electrophoresis indicates that p12*, the neck appendage protein, is transferred to the particles when complex is formed. Complex can also be produced by incubation of 12−13− particles at 42°, by incubation at pH 6.0, or by incubation in the presence of 20 mM EDTA. Complex is also formed from DNA-containing 12− particles but to a lesser extent.
Publisher version (URL):http://dx.doi.org/10.1016/0042-6822(81)90347-0
URI:http://hdl.handle.net/10261/39090
ISSN:0042-6822
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