Separation and quantitation of intracellular forms of poliovirus RNA by agarose gel electrophoresis.

Abstract:

:Intracellular poliovirus-specific RNA species can be measured directly by electrophoresis of total cytoplasmic nucleic acids through 1% agarose gels, resulting in the separation of single- and double-stranded forms of poliovirus RNA from each other and from HeLa cell 28S ribosomal RNA. Single-stranded RNA molecules differing by only 15% in length are resolved in this gel system. RNA species can be visualized as fluorescen bands appearing after staining of the gels with ethidium bromide and observation under ultraviolet illumination. The total amount of RNA can be determined by densitometric quantitation of the fluorescent response. In this way, the amount of poliovirus-specific RNA within the cytoplasm of HeLa cells infected for various times has been estimated. At 170-min postinfection, there are 0.67 X 10(5) molecules of single-stranded poliovirus RNA per cell and at 230 min, the amount has increased to 3.7 X 10(5) molecules/cell. Poliovirus double-strnaded RNA reaches a maximum of 0.7 X 10(5) molecules/cell at 330 min after infection.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Hewlett MJ,Rozenblatt S,Ambros V,Baltimore D

doi

10.1021/bi00631a027

subject

Has Abstract

pub_date

1977-06-14 00:00:00

pages

2763-7

issue

12

eissn

0006-2960

issn

1520-4995

journal_volume

16

pub_type

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