Chapter title |
A Molecular Toolbox to Engineer Site-Specific DNA Replication Perturbation
|
---|---|
Chapter number | 20 |
Book title |
Genome Instability
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7306-4_20 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7305-7, 978-1-4939-7306-4
|
Authors |
Nicolai B. Larsen, Ian D. Hickson, Hocine W. Mankouri |
Abstract |
Site-specific arrest of DNA replication is a useful tool for analyzing cellular responses to DNA replication perturbation. The E. coli Tus-Ter replication barrier can be reconstituted in eukaryotic cells as a system to engineer an unscheduled collision between a replication fork and an "alien" impediment to DNA replication. To further develop this system as a versatile tool, we describe a set of reagents and a detailed protocol that can be used to engineer Tus-Ter barriers into any locus in the budding yeast genome. Because the Tus-Ter complex is a bipartite system with intrinsic DNA replication-blocking activity, the reagents and protocols developed and validated in yeast could also be optimized to engineer site-specific replication fork barriers into other eukaryotic cell types. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 9 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 3 | 33% |
Researcher | 3 | 33% |
Student > Bachelor | 1 | 11% |
Other | 1 | 11% |
Professor > Associate Professor | 1 | 11% |
Other | 0 | 0% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 6 | 67% |
Nursing and Health Professions | 1 | 11% |
Agricultural and Biological Sciences | 1 | 11% |
Unknown | 1 | 11% |