Canadian Forest Service Publications

Preferential separation of cytoplasmic-polyhedrosis virus (CPV) RNAs from infected midgut cells. 1972. Richards, W.C.; Hayashi, Y. Journal of Invertebrate Pathology 20:200-207.

Year: 1972

Issued by: Great Lakes Forestry Centre

Catalog ID: 36662

Language: English

Availability: PDF (request by e-mail)

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Viral progeny, double-stranded RNA (dsRNA), and virus-specific single-stranded RNA (ssRNA) synthesized in midgut cells of Malacosoma disstria larvae infected with cytoplasmic-polyhedrosis virus (CPV), have been separated using Mg2+ for RNA aggregation, and lithium chloride for differential solubilization between the ds- and ssRNA.

Under high concentrations of Mg2+ (5 × 10−3 and 10−2 m MgSO4), dsRNA was not affected, remaining positional at a sedimentation rate of 15S, but virus-specific ssRNA and host ribosomal RNA as single-stranded (rRNA), were separated from dsRNA by aggregation. After RNase treatment there was no appreciable contamination of virus specific ssRNA and rRNA.

It was found that ds and ssRNA could be separated in 2 mLiCl on the basis of their different solubilities. The ssRNA fraction obtained by such a procedure contained only 6% dsRNA, and the dsRNA fraction was free of ssRNA.

Further separation of dsRNA by both the Mg2+ and LiCl treatments was accomplished through prolonged centrifugation which yielded 12 and 15 S components; these are the same characteristics found for viral genome RNA appearing in the sedimentation profile.