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[Isolation and translation of late mRNA coding for the structural proteins of simian adenovirus SA7]
Abstract:
A total preparation of cytoplasmic RNA was isolated from late stage-infected African green monkey kidney cells using phenol extraction procedure. The poly (A)-containing mRNA fraction was selected on oligo(dT)-cellulose columns. The resulting mRNA preparations were heterogenous in size and contained about 20--60% of SA7-derived sequences. SA7 late mRNA was efficiently translated in rabbit reticulocyte cell-free system giving rise to a number of polypeptide products that were related by antigenicity to authentic SA7 virion proteins. The main translation product having a molecular weight of 114 kilodaltons was identified as intact SA7 hexon protein.
Insights
Researchers isolated African green monkey kidney cell RNA to study SA7 virus. They identified the main translation product as the 114-kilodalton SA7 hexon protein using a cell-free system.
Area of Science:
- Molecular Biology
- Virology
- Cell Biology
Background:
- African green monkey kidney cells are a common model for studying viral replication.
- Simian Adenovirus 7 (SA7) is a virus that infects primate cells.
- Understanding viral gene expression is crucial for developing antiviral strategies.
Purpose of the Study:
- To isolate and characterize late-stage mRNA from SA7-infected cells.
- To investigate the translation products of SA7 late mRNA.
- To identify specific viral proteins synthesized during SA7 infection.
Main Methods:
- Cytoplasmic RNA isolation using phenol extraction.
- Polyadenylated mRNA selection via oligo(dT)-cellulose chromatography.
- In vitro translation using a rabbit reticulocyte cell-free system.
- Antigenic analysis and molecular weight determination of translation products.
Main Results:
- Heterogeneous mRNA preparations containing 20-60% SA7-derived sequences were obtained.
- SA7 late mRNA efficiently translated into multiple polypeptide products.
- These products showed antigenic similarity to authentic SA7 virion proteins.
- The primary translation product, a 114-kilodalton polypeptide, was identified as the intact SA7 hexon protein.
Conclusions:
- SA7 late mRNA encodes viral proteins, including the hexon protein.
- The cell-free translation system is effective for studying SA7 gene expression.
- This study contributes to the understanding of SA7 virus replication and protein synthesis.