| Chapter title |
Proteoglycans: Gene Cloning
|
|---|---|
| Chapter number | 1 |
| Book title |
Proteoglycans
|
| Published in |
Methods in molecular biology, January 2012
|
| DOI | 10.1007/978-1-61779-498-8_1 |
| Pubmed ID | |
| Book ISBNs |
978-1-61779-497-1, 978-1-61779-498-8
|
| Authors |
Mauricio Cortes, James R. Mensch, Miriam Domowicz, Nancy B. Schwartz, Cortes, Mauricio, Mensch, James R., Domowicz, Miriam, Schwartz, Nancy B. |
| Abstract |
Aggrecan is a large proteoglycan that plays roles in numerous tissues during vertebrate development and adult life. The 6,327-nt chick aggrecan coding sequence had been determined from overlapping clones, but a full-length cDNA, needed for use in transgenic expression studies, had not been constructed. The strategy employed to do so was to generate two overlapping cDNA subfragments that shared a unique restriction site in the overlap and then join them at that site. These subfragments were obtained and cloned into the TOPO-TA vector pCR2.1. Digestion of the two constructs with the shared-site enzyme, XbaI, produced vector/5'-cDNA and 3'-cDNA fragments with XbaI-ends; these were ligated to produce the final full-length cDNA. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 4 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Professor > Associate Professor | 1 | 25% |
| Researcher | 1 | 25% |
| Unknown | 2 | 50% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 1 | 25% |
| Neuroscience | 1 | 25% |
| Unknown | 2 | 50% |