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  • Title: Morphogenesis of phi X174: in vitro synthesis of infectious phage from purified viral components.
    Author: Aoyama A, Hamatake RK, Hayashi M.
    Journal: Proc Natl Acad Sci U S A; 1981 Dec; 78(12):7285-9. PubMed ID: 6461002.
    Abstract:
    An in vitro system that synthesizes infectious phage phi X174 was developed. The synthesis depended on phi X174 supercoiled replicative form DNA, purified phi X174 gene A protein, gene C protein, gene J protein, prohead (phage head precursor composed of gene F, G, H, B, and D proteins), and uninfected host crude extract. The infectious phage synthesis was coupled with DNA synthesis. De novo initiation, elongation, and termination of phi X174 single-stranded DNA was observed. The phage synthesized in vitro cosedimented with in vivo phage in sucrose gradients and had the same buoyant density as in vivo phage in a CsCl gradient. Our results indicate that the in vitro system mimics the in vivo phi X174 assembly process.
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