Cataloging altered gene expression in young and senescent cells using enhanced differential display
| Autor: | Maarten H.K. Linskens, Shahin M. Saati, William H. Andrews, Bryant Villeponteau, Junli Feng, Walter Funk, Brett E. Enlow, Leath A. Tonkin |
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| Přispěvatelé: | Groningen Biomolecular Sciences and Biotechnology |
| Rok vydání: | 1995 |
| Předmět: |
Cloning
Differential display Base Sequence Hybridization probe Molecular Sequence Data Gene Expression Fibroblasts Biology Polymerase Chain Reaction Molecular biology Gene expression Genetics Humans RNA Messenger Northern blot Differential display technique DNA Probes Gene Cell aging Cells Cultured Cellular Senescence DNA Primers |
| Zdroj: | Nucleic Acids Research, 23(16). Oxford University Press |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/23.16.3244 |
| Popis: | Recently, a novel PCR-based technique, differential display (DD), has facilitated the study of differentially expressed genes at the mRNA level. We report here an improved version of DD, which we call Enhanced Differential Display (EDD). We have modified the technique to enhance reproducibility and to facilitate sequencing and cloning. Using EDD, we have generated and verified a catalog of genes that are differentially expressed between young and senescent human diploid fibroblasts (HDF). From 168 genetags that were identified initially, 84 could be sequenced directly from PCR amplified bands. These sequences represent 27 known genes and 37 novel genes. By Northern blot analysis we have confirmed the differential expression of a total of 23 genes (12 known, 11 novel), while 19 (seven known, 12 novel) did not show differential expression. Several of the known genes were previously observed by others to be differentially expressed between young and senescent fibroblasts, thereby validating the technique. |
| Databáze: | OpenAIRE |
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