| Chapter title |
Detection of Separase Activity Using a Cleavage Sensor in Live Mouse Oocytes
|
|---|---|
| Chapter number | 11 |
| Book title |
Mouse Oocyte Development
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-8603-3_11 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-8602-6, 978-1-4939-8603-3
|
| Authors |
Elvira Nikalayevich, Nora Bouftas, Katja Wassmann, Nikalayevich, Elvira, Bouftas, Nora, Wassmann, Katja |
| Abstract |
Separase proteolytically removes cohesin complexes from sister chromatid arms in meiosis I, which is essential for chromosome segregation. Regulation of separase activity is essential for proper cell cycle progression and correct chromosome segregation. Onset of endogenous separase activity has not yet been observed in live oocytes.We describe here a method for detecting separase activity in mouse oocytes in vivo. This method utilizes a previously described cleavage sensor made up of H2B-mCherry fused with Scc1(107-268 aa)-YFP. The cleavage sensor is loaded on the chromosomes through its H2B-tag, and the signal from both mCherry and YFP is visible. Upon separase activation the Scc1 fragment is cleaved and YFP dissociates from the chromosomes. The change in the ratio between mCherry and YFP fluorescence intensity is a readout of separase activity. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 2 | 25% |
| Sweden | 1 | 13% |
| India | 1 | 13% |
| Japan | 1 | 13% |
| Unknown | 3 | 38% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 4 | 50% |
| Members of the public | 4 | 50% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 22 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 4 | 18% |
| Student > Master | 4 | 18% |
| Student > Ph. D. Student | 2 | 9% |
| Student > Doctoral Student | 1 | 5% |
| Other | 1 | 5% |
| Other | 2 | 9% |
| Unknown | 8 | 36% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 7 | 32% |
| Agricultural and Biological Sciences | 5 | 23% |
| Veterinary Science and Veterinary Medicine | 1 | 5% |
| Engineering | 1 | 5% |
| Unknown | 8 | 36% |