Rapid and efficient purification of Cowpea chlorotic mottle virus by sucrose cushion ultracentrifugation

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

A simple, economical and efficient method for isolation of Cowpea chlorotic mottle virus (CCMV) particles was developed. Abundant viral particles could be obtained with three steps of 10 min duration each of conventional differential centrifugations and one sucrose cushion ultracentrifugation step of 2 h. Characterization of purified virus assessed by transmission electron microscopy, polyacrylamide gel electrophoresis (PAGE) and Western blot revealed the typical isometric particles of CCMV without any visible contamination or degradation. RNA extracted from purified CCMV particles showed all four RNA components by agarose gel electrophoresis. Both purified virus particles and RNA were biologically active and initiated successful infection upon inoculation to cowpea.

Original languageEnglish (US)
Pages (from-to)84-86
Number of pages3
JournalJournal of Virological Methods
Volume141
Issue number1
DOIs
StatePublished - Apr 1 2007

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Bromovirus
Ultracentrifugation
Virion
Sucrose
RNA
Agar Gel Electrophoresis
Transmission Electron Microscopy
Centrifugation
Polyacrylamide Gel Electrophoresis
Western Blotting
Viruses
Infection

All Science Journal Classification (ASJC) codes

  • Virology

Cite this

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abstract = "A simple, economical and efficient method for isolation of Cowpea chlorotic mottle virus (CCMV) particles was developed. Abundant viral particles could be obtained with three steps of 10 min duration each of conventional differential centrifugations and one sucrose cushion ultracentrifugation step of 2 h. Characterization of purified virus assessed by transmission electron microscopy, polyacrylamide gel electrophoresis (PAGE) and Western blot revealed the typical isometric particles of CCMV without any visible contamination or degradation. RNA extracted from purified CCMV particles showed all four RNA components by agarose gel electrophoresis. Both purified virus particles and RNA were biologically active and initiated successful infection upon inoculation to cowpea.",
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Rapid and efficient purification of Cowpea chlorotic mottle virus by sucrose cushion ultracentrifugation. / Ali, Akhtar; Roossinck, Marilyn J.

In: Journal of Virological Methods, Vol. 141, No. 1, 01.04.2007, p. 84-86.

Research output: Contribution to journalArticle

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