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Double-stranded Ribonucleic Acid from Cytoplasmic Polyhedrosis Virus of the Silkworm
1Nagoya University, Chikusa, Nagoya, and University of Tokyo, Hongo, Tokyo, Japan.
Abstract:
Ribonucleic acid (RNA) was extracted by phenol treatment from cytoplasmic polyhedrosis virus isolated from the midgut of infected silkworms. This RNA appears as threads when precipitated in alcohol. Two components having different sedimentation constants were observed. The molecular weight of the RNA preparation obtained by sedimentation coefficient (weight-averaged) and intrinsic viscosity was about 2 x 10(6) to 3 x 10(6). It was one-half to one-third the size of the calculated molecular weight for an entire RNA molecule in a virion. Electron micrographs of this RNA preparation showed two peaks in the distribution of contour length, at 0.4 and 1.3 mum, which would correspond to molecular weights of 10(6) and 3 x 10(6), respectively. The extracted RNA seemed to split into segments at a preferential breaking point. This RNA was soluble in concentrated salt solution, differing from single stranded high-molecular-weight RNA. The base composition of this RNA was complementary in the ratios of adenosine to uridine and guanosine to cytosine. It contained 43% guanosine plus cytosine. Based on its filamentous appearance by electron microscopy, typical pattern of optical rotatory dispersion and circular dichroism, sharp transition of the optical properties on heating, great hyperchromicity on degradation, nonreactivity with formaldehyde, and resistance to ribonucleases, it is concluded that this RNA is double-stranded and has regular base pairings of guanosine-cytosine and adenosine-uridine.
Insights
Researchers extracted double-stranded ribonucleic acid (RNA) from silkworm cytoplasmic polyhedrosis virus. This viral RNA, smaller than expected, appears segmented and exhibits characteristics of double-stranded nucleic acids.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Cytoplasmic polyhedrosis virus (CPV) infects silkworms, impacting silk production.
- Understanding viral RNA structure is crucial for virology and molecular biology.
Purpose of the Study:
- To characterize the ribonucleic acid (RNA) extracted from CPV.
- To determine the structural properties and molecular weight of the viral RNA.
Main Methods:
- Phenol extraction of RNA from CPV.
- Sedimentation analysis and intrinsic viscosity measurements.
- Electron microscopy for contour length distribution.
- Spectroscopic analysis (optical rotatory dispersion, circular dichroism) and thermal denaturation.
- Biochemical assays (ribonuclease resistance, formaldehyde reactivity).
Main Results:
- Extracted RNA appeared as threads and showed two sedimentation components.
- Molecular weight estimations ranged from 2-3 x 10^6, smaller than expected for a full genome.
- Electron microscopy revealed segmented RNA with molecular weights of 10^6 and 3 x 10^6.
- RNA exhibited properties indicative of double-stranded structure, including complementary base ratios and resistance to degradation.
Conclusions:
- The extracted CPV RNA is double-stranded.
- The RNA molecule appears to be segmented, with preferential breaking points.
- The observed properties align with regular base pairing (G-C, A-U).
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