Construction of a microphage variant of filamentous bacteriophage.
The intergenic region in the genome of the Ff class of filamentous phage (comprising strains fl, fd and M13) genome constitutes 8% of the viral genome, and has essential functions in DNA replication and phage morphogenesis. The functional domains of this region may be inserted into separate sites of a plasmid to function independently. Here, we demonstrate the construction of a plasmid containing, sequentially, the origin of (+)-strand synthesis, the packaging signal and a terminator of (+)-strand synthesis. When host cells harboring this plasmid (pLS7) are infected with helper phage they produce a microphage particle containing all the structural elements of the mature, native phage. The microphage is 65 A in diameter and about 500 A long. It contains a 221-base single-stranded circle of DNA coated by about 95 copies of the major coat protein (gene 8 protein).
Specthrie
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Department of Biochemistry and Molecular Biophysics, College of Physicians and Surgeons, Columbia University, New York, NY 10032.
Bullitt
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Horiuchi
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Model
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Russel
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M
Makowski
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L
D017104:Bacteriophage M13 / Q000235:genetics* / Q000648:ultrastructure; D001483:Base Sequence; D003090:Coliphages / Q000235:genetics* / Q000648:ultrastructure; D004261:DNA Replication; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000235:genetics*; D015967:Gene Expression Regulation, Viral; D006378:Helper Viruses / Q000235:genetics; D008969:Molecular Sequence Data; D009024:Morphogenesis; D009690:Nucleic Acid Conformation; D010316:Particle Size; D010957:Plasmids / Q000235:genetics* / Q000648:ultrastructure
D004274:DNA, Recombinant; D004279:DNA, Viral