Fractionation of Corynebacterium pekinense AS 1.299 phage subtypes by anion-exchange chromatography

G.R. Chen, X.H. Xu, W.H. Deng, W.H. Lu, R.M. Chen, R.Z. Zhang, L. Li, Tianbao Chen, X.Y. Lin, P.F. Rao

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

A novel method to fractionate phage into its subtypes while fully retaining biological function is reported. Corynebacterium pekinense AS 1.299 phage samples, purified by either conventional ultracentrifugation or gel chromatography on a Superose® 6 Prep column (0.78×30 cm), were fractionated further into four fractions by anion-exchange chromatography on a Toyopearl SuperQ 650C column (0.5×20 cm) with a linear gradient of NaCl concentration from 0.2 to 1.0 M in 0.02 M carbonate–biocarbonate buffer, pH 10.0. Two peaks were identified to be C. pekinense AS 1.299 phages by their ability to infect the host bacteria when inoculated into the culture media, and when examined by electron microscopy. These two types of the phage were found to be morphologically the same except for the difference in the length of their non-contractile tails. Both possessed an isometric head with a diameter of 50±3 nm, while their tails were 170±10 and 210±10 nm, respectively. This simple technique provides a convenient method for phage isolation not only to its species homogeneity, but also to determine its subtype or variant homogeneity.
Original languageEnglish
Pages (from-to)11-17
Number of pages7
JournalJournal of Virological Methods
Volume95(1-2)
Issue number1-2
DOIs
Publication statusPublished - Jun 2001

ASJC Scopus subject areas

  • Virology

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