| Autor: | V. Lunyak, L. Noskin, N. Giliano, R. Burgess |
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| Rok vydání: | 2003 |
| Předmět: | |
| Zdroj: | Molecular Biology. 37:140-147 |
| ISSN: | 0026-8933 |
| Popis: | A modified version of the chromosomal immunoprecipitation (ChIP) assay was implemented for discrete isolation and characterization of actively transcribed genes. Specifically, it was demonstrated with the gene II/9-1 of Sciara coprophila as a model locus that significant enhancement in the isolation of actively transcribed versus repressed and inactive genes can be achieved through the ChIP methodology. A combination of solid-phase magnetic bead technology with chromosomal immunoprecipitation using antibodies that recognize the large subunit (c) of RNA polymerase II resulted in efficient isolation of the promoter region of gene II/9-1 exclusively during the amplification stage of larval development, when the gene is actively transcribed. It is postulated that the novel technology described herein can be applied to a wide variety of systems for efficient isolation and in vivo assessment of actively transcribed genes regulated by virtually any given transcription factor. |
| Databáze: | OpenAIRE |
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