| Chapter title |
Chromatin structure analysis of single gene molecules by psoralen cross-linking and electron microscopy.
|
|---|---|
| Chapter number | 9 |
| Book title |
The Nucleus
|
| Published in |
Methods in molecular biology, January 2015
|
| DOI | 10.1007/978-1-4939-1680-1_9 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-1679-5, 978-1-4939-1680-1
|
| Authors |
Christopher R Brown, Julian A Eskin, Stephan Hamperl, Joachim Griesenbeck, Melissa S Jurica, Hinrich Boeger, Christopher R. Brown, Julian A. Eskin, Melissa S. Jurica, Brown, Christopher R., Eskin, Julian A., Hamperl, Stephan, Griesenbeck, Joachim, Jurica, Melissa S., Boeger, Hinrich |
| Abstract |
Nucleosomes occupy a central role in regulating eukaryotic gene expression by blocking access of transcription factors to their target sites on chromosomal DNA. Analysis of chromatin structure and function has mostly been performed by probing DNA accessibility with endonucleases. Such experiments average over large numbers of molecules of the same gene, and more recently, over entire genomes. However, both digestion and averaging erase the structural variation between molecules indicative of dynamic behavior, which must be reconstructed for any theory of regulation. Solution of this problem requires the structural analysis of single gene molecules. In this chapter, we describe a method by which single gene molecules are purified from the yeast Saccharomyces cerevisiae and cross-linked with psoralen, allowing the determination of nucleosome configurations by transmission electron microscopy. We also provide custom analysis software that semi-automates the analysis of micrograph data. This single-gene technique enables detailed examination of chromatin structure at any genomic locus in yeast. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United Kingdom | 1 | 33% |
| United States | 1 | 33% |
| Unknown | 1 | 33% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 2 | 67% |
| Scientists | 1 | 33% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 19 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 5 | 26% |
| Researcher | 4 | 21% |
| Unspecified | 1 | 5% |
| Professor | 1 | 5% |
| Lecturer | 1 | 5% |
| Other | 2 | 11% |
| Unknown | 5 | 26% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 6 | 32% |
| Agricultural and Biological Sciences | 4 | 21% |
| Chemistry | 2 | 11% |
| Unspecified | 1 | 5% |
| Neuroscience | 1 | 5% |
| Other | 1 | 5% |
| Unknown | 4 | 21% |