| Chapter title |
Electrophoretic Analysis of the DNA Supercoiling Activity of DNA Gyrase
|
|---|---|
| Chapter number | 15 |
| Book title |
Molecular Motors
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-8556-2_15 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-8554-8, 978-1-4939-8556-2
|
| Authors |
Belén Martínez-García, Antonio Valdés, Joana Segura, Silvia Dyson, Ofelia Díaz-Ingelmo, Joaquim Roca, Martínez-García, Belén, Valdés, Antonio, Segura, Joana, Dyson, Silvia, Díaz-Ingelmo, Ofelia, Roca, Joaquim |
| Abstract |
Most bacterial cells have a motor enzyme termed DNA gyrase, which is a type-2 topoisomerase that reduces the linking number (Lk) of DNA. The supercoiling energy generated by gyrase is essential to maintain the bacterial chromosome architecture and regulate its DNA transactions. This chapter describes the use of agarose-gel electrophoresis to detect the unconstrained supercoiling of DNA generated by gyrase or other gyrase-like activities. Particular emphasis is made on the preparation of a relaxed plasmid as initial DNA substrate, on the distinction of constrained and unconstrained DNA supercoils, and on the measurement of the DNA supercoiling density achieved by gyrase activity. |
X Demographics
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Demographic breakdown
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Mendeley readers
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|---|---|---|
| Unknown | 11 | 100% |
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| Researcher | 2 | 18% |
| Professor | 1 | 9% |
| Other | 1 | 9% |
| Student > Master | 1 | 9% |
| Other | 1 | 9% |
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