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Abstract:
Techniques are described for preparing intact Simian Hemorrhagic Fever (SHF) virus RNA. SHF RNA extracted by proteinase K digestion in the presence of sodium dodecyl sulphate (SDS) has a sedimentation coefficient of 49 S compared with a reference figure of 47 S for Sindbis RNA. Purified SHF RNA in cesium sulphate gradient has a buoyant density of 1.63 g/ml similar to that of Sindbis RNA. This result leads to the conclusion that SHF RNA is single stranded. This is supported by results on RNAse sensitivity and analyses on sucrose gradients under different ionic strength conditions. Electrophoresis analyses on both hydroxymethylmercuric acid or formaldehyde containing gels, gave a value of about 5.5 X 10(6) daltons for the molecular weight of SHF RNA. No evidence of subunit structure was found. From these results, we conclude that the SHF virus genome is a single continuous chain of about 15,000 ribonucleotides.
Insights
Researchers characterized Simian Hemorrhagic Fever (SHF) virus RNA, determining it is a single-stranded genome. This single, continuous RNA chain has an estimated molecular weight of 5.5 million daltons.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Simian Hemorrhagic Fever (SHF) virus is an important pathogen.
- Understanding viral genome structure is crucial for studying viral replication and pathogenesis.
Purpose of the Study:
- To characterize the Simian Hemorrhagic Fever virus RNA.
- To determine the physical and chemical properties of SHF virus RNA.
Main Methods:
- RNA extraction using proteinase K and sodium dodecyl sulphate (SDS).
- Sedimentation coefficient analysis.
- Buoyant density centrifugation in cesium sulphate gradients.
- RNAse sensitivity assays.
- Sucrose gradient analysis under varying ionic strengths.
- Electrophoresis on hydroxymethylmercuric acid or formaldehyde gels.
Main Results:
- SHF virus RNA exhibited a sedimentation coefficient of 49 S.
- Buoyant density was determined to be 1.63 g/ml, indicating single-stranded RNA.
- RNAse sensitivity and ionic strength analyses supported the single-stranded nature.
- Molecular weight was estimated at approximately 5.5 x 10^6 daltons.
- No evidence of subunit structure was observed.
Conclusions:
- The Simian Hemorrhagic Fever virus genome is composed of a single, continuous, single-stranded RNA molecule.
- The estimated size of the SHF virus RNA genome is approximately 15,000 ribonucleotides.