Selective separation of virus proteins and double-stranded RNAs by SDS-KCl precipitation

Joseph K K Li, Todd Johnson, Yi Yuan Yang, Vicki Shore

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The total viral structural polypeptides and the double-stranded genomic RNAs of bluetongue virus can be selectively separated by a single SDS-KCl precipitation step. This simple, rapid and highly reproducible method enables greater than 95% recovery and purity of both viral proteins and dsRNAs within 30 min. The serotypic identity of the separated dsRNAs can be analyzed by SDS-PAGE electrophorogram immediately. After a single phenol/chloroform extraction, the dsRNA can also be used as hybridization probes, templates for molecular cloning and direct RNA sequencing. The SDS-KCl-precipitated viral proteins could be used readily for peptide mapping and as immunogens. Polyclonal and monoclonal antibodies raised against SDS-KCl-precipitated viral structural polypeptides were useful in Western immunoblots.

Original languageEnglish
Pages (from-to)3-15
Number of pages13
JournalJournal of Virological Methods
Volume26
Issue number1
DOIs
Publication statusPublished - 1989
Externally publishedYes

Fingerprint

Double-Stranded RNA
Viral Proteins
Bluetongue virus
Viruses
RNA Sequence Analysis
Peptides
Peptide Mapping
RNA Viruses
Molecular Cloning
Chloroform
Phenol
Polyacrylamide Gel Electrophoresis
Proteins
Western Blotting
Monoclonal Antibodies

Keywords

  • dsRNA purification
  • Protein isolation
  • SDS-KCl precipitation

ASJC Scopus subject areas

  • Virology

Cite this

Selective separation of virus proteins and double-stranded RNAs by SDS-KCl precipitation. / Li, Joseph K K; Johnson, Todd; Yang, Yi Yuan; Shore, Vicki.

In: Journal of Virological Methods, Vol. 26, No. 1, 1989, p. 3-15.

Research output: Contribution to journalArticle

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