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Updated: Jul 7, 2026
05:22
Electrophoretic Analysis of Replication Through Structure-Prone DNA Repeats Within the SV40-Based Human Episome
Published on: September 13, 2024
In vitro analysis of SV40 DNA replication
Current Protocols in Cell Biology
|January 30, 2008
Summary
This study details a method to investigate cellular factors influencing eukaryotic DNA replication. It utilizes a specialized plasmid and viral helicase to analyze replication products and purification protocols.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Eukaryotic DNA replication is a complex process involving numerous cellular factors.
- Understanding these factors is crucial for comprehending cell division and disease.
Purpose of the Study:
- To describe a method for studying cellular factors that impact eukaryotic replication.
- To provide protocols for the expression, purification, and analysis of key replication components.
Main Methods:
- Utilizing a plasmid with a Simian virus 40 (SV40) origin of replication.
- Supplying the viral helicase, T antigen, and replication factors from S100 extracts of 293 cells.
- Performing analysis of replication products.
Main Results:
- The described method allows for the investigation of cellular contributions to DNA replication.
- Protocols for T antigen purification and replication product analysis are provided.
Conclusions:
- This unit offers a robust methodology for dissecting eukaryotic DNA replication mechanisms.
- The provided protocols facilitate further research into replication factor function.
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Two states at the origin of replication
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