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BACTERIAL VIRUSES

Mechanism of Adsorption and Eclipse of Bacteriophage φX174 II. Attachment and Eclipse with Isolated Escherichia coli Cell Wall Lipopolysaccharide

Nino L. Incardona, LeRoy Selvidge
Nino L. Incardona
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LeRoy Selvidge
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ABSTRACT

A mixture of aqueous phenol, choloroform, and ether extracts the lipopolysaccharides (LPS) from the φX174-sensitive strain, Escherichia coli C/1, and resistant strains, C/φX and K12. Interaction of the C/1 LPS with φX in a starvation buffer containing 10−3 M CaCl2 at 37 C, but not at 15 C, results in a first-order inactivation that is specific for C/1 LPS. After interaction for 60 min at 15 C, followed by centrifugation, 37 and 20% of a 14C-φX preparation are bound to the C/1 and C/φX LPS pellets, respectively. The results for intact cells are 75 and 10%. Supporting the conclusion that this represents specific attachment of φX to its receptor site in the LPS is the fact that EDTA-borate buffer is required to elute 85% of the 14C-φX from the C/1 LPS, whereas starvation buffer elutes the same amount from C/φX LPS. Moreover, 95% of the PFU are found in the C/1 LPS pellets as compared with 50% in the resistant strain LPS pellets. When the products of interaction between φX and LPS at 37 C are examined by sucrose density gradients in EDTA-borate, a single 60 to 90S peak is observed in the C/1 sample, and the single peak cosediments with the 120S marker φX in the C/φX sample. This change in S20, w is very similar to that reported for the eclipse of φX in vivo. If the inactivation at 37 C is carried out on φX-LPS complexes first formed at 15 C, the first-order kinetics are biphasic and nearly identical to that observed for the eclipse kinetics of φX attached to intact cells. Thus, the φX-LPS system is suitable for in vitro studies on the early events in φX infection.

  • Copyright © 1973 American Society for Microbiology
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Mechanism of Adsorption and Eclipse of Bacteriophage φX174 II. Attachment and Eclipse with Isolated Escherichia coli Cell Wall Lipopolysaccharide
Nino L. Incardona, LeRoy Selvidge
Journal of Virology May 1973, 11 (5) 775-782; DOI:

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Mechanism of Adsorption and Eclipse of Bacteriophage φX174 II. Attachment and Eclipse with Isolated Escherichia coli Cell Wall Lipopolysaccharide
Nino L. Incardona, LeRoy Selvidge
Journal of Virology May 1973, 11 (5) 775-782; DOI:
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