A Flexible Approach to Studying Post-Transcriptional Gene Regulation in Stably Transfected Mammalian Cells
| Autor: | B. JoNell Hamilton, John Botson, Ralph C. Nichols, Xiao Wei Wang, William F. C. Rigby, Jane E. Collins, Victoria Uribe, Seth A. Brooks, Moe Zan |
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| Rok vydání: | 2010 |
| Předmět: |
Green Fluorescent Proteins
Molecular Sequence Data Bioengineering Biology Applied Microbiology and Biotechnology Biochemistry Article Plasmid Transcription (biology) Complementary DNA Humans RNA Messenger Regulatory Elements Transcriptional 3' Untranslated Regions Molecular Biology Gene Regulation of gene expression Reporter gene Base Sequence Models Genetic Tumor Necrosis Factor-alpha HEK 293 cells Transfection Flow Cytometry Molecular biology HEK293 Cells Gene Expression Regulation Data Interpretation Statistical Biotechnology |
| Zdroj: | Molecular Biotechnology. 48:210-217 |
| ISSN: | 1559-0305 1073-6085 |
| DOI: | 10.1007/s12033-010-9360-8 |
| Popis: | The study of post-transcriptional regulation is constrained by the technical limitations associated with both transient and stable transfection of chimeric reporter plasmids examining the activity of 3′-UTR cis-acting elements. We report the adaptation of a commercially available system that enables consistent stable integration of chimeric reporter cDNA into a single genomic site in which transcription is induced by tetracycline. Using this system, we demonstrate the tight control afforded by this system and its suitability in mapping the regulatory function of defined cis-acting elements in the human TNF 3′-UTR, as well as the distinct effects of serum starvation on transiently transfected and stably integrated chimeric reporter genes. |
| Databáze: | OpenAIRE |
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