PMID: 1202245Dec 1, 1975Paper

Purification of influenza viral complementary RNA: its genetic content and activity in wheat germ cell-free extracts

Journal of Virology
P R Etkind, R M Krug

Abstract

Influenza viral complementary RNA (cRNA) was purified free from any detectable virion-type RNA (vRNA), and its genetic content and activity in wheat germ cell-free extracts were examined. After phenol-chloroform extraction of cytoplasmic fractions from infected cells, poly(A)-containing viral cRNA is found in two forms: in single-stranded RNA and associated with vRNA in partially and fully double-stranded RNA. To purify single-stranded cRNA free of these double-stranded forms, it was necessary to employ, as starting material, RNA fractions in which cRNA was predominantly single stranded. Two RNA fractions were successfully employed as starting material: polyribosomal RNA and the total cytoplasmic RNA from infected cells treated with 100 mug of cycloheximide (CM) per ml at 3 h after infection. In WSN virus-infected canine kidney (MDCK) cells, the addition of CM at 3 h after infection stimulates the production of cRNA threefold and causes a very large increase in the proportion of the cytoplasmic cRNA which is single stranded; double-stranded RNA forms are greatly reduced in amount. Total cRNA was obtained by oligo(dT)-cellulose chromatography, and single-stranded cRNA was separated from double-stranded forms by Sepharose 4B chro...Continue Reading

References

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Citations

Apr 17, 1980·Nature·W J BeanA P Kendal
May 1, 1976·Journal of Virology·J Content
May 1, 1976·Journal of Virology·P Tekamp, E E Penhoet
Nov 1, 1979·Journal of Virology·J SpragueJ B Milstien
Dec 1, 1980·Journal of Virology·D M LambertK Dorsch-Hasler
May 1, 1977·Journal of Virology·J ContentM Horisberger
May 1, 1977·Journal of Virology·P R EtkindR M Krug
Oct 1, 1985·Journal of Virology·S NakadaP Palese
Oct 1, 1984·Molecular and Cellular Biology·M G Katze, R M Krug
Feb 1, 1978·Journal of Virology·S J Plotch, R M Krug
Jul 15, 1976·Virology·M B Richey, P Palese
Nov 1, 1978·Virology·R A LambP W Choppin
Oct 1, 1976·Journal of Virology·D P NayakF O Wettstein

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