Recombinant plant dsRNA-binding protein as an effective tool for the isolation of viral replicative form dsRNA and universal detection of RNA viruses

Technique

Abstract

The isolation of viral replicative form (RF) double-stranded RNA (dsRNA) is a classic technique for plant virus detection when the virus species cannot be predicted from disease symptoms. However, the method has not been very widely used, most likely because dsRNA isolation using CF-11 cellulose is laborious and time-consuming. Here we report an alternative tool, a recombinant plant dsRNA-binding protein, to isolate dsRNA. This tool enables us to isolate viral RF dsRNA in an hour from either extracted nucleic acids or crude detergent extracts. Combining this technique with sequence-non-specific reverse transcription, PCR amplification, cloning, and sequencing, a variety of viruses were efficiently detected using a single set of reagents and procedures.

Keywords

Detection dsRNA dsRNA-binding protein Replicative form RNA virus 

Supplementary material

10327_2009_155_MOESM1_ESM.doc (590 kb)
Supplementary material 1 (DOC 590 kb)
10327_2009_155_MOESM2_ESM.doc (35 kb)
Supplementary material 2 (DOC 35 kb)
10327_2009_155_MOESM3_ESM.doc (32 kb)
Supplementary material 3 (DOC 31 kb)

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Copyright information

© The Phytopathological Society of Japan and Springer 2009

Authors and Affiliations

  • Kappei Kobayashi
    • 1
  • Reiko Tomita
    • 1
  • Masaru Sakamoto
    • 1
  1. 1.Iwate Biotechnology Research CenterKitakamiJapan

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