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DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract.
Open accessAbstract
We have developed a cell-free system for studying the synthesis of mRNA in mammalian cells. The system consists of a dialyzed and concentrated whole-cell extract derived from HeLa cells, small molecules and cofactors needed for transcription, and exogenously added DNA. Accurate transcription by RNA polymerase II is entirely dependent upon addition of promoter-containing eukaryotic DNA. At optimal DNA and extract concentrations, transcription initiation from the adenovirus serotype 2 late promoter is readily detectable, and specific transcripts over 4000 nucleotides in length are observed. The RNA synthesized in vitro contains the same 5' capped RNase T1 undecanucleotide as does the in vivo transcript. RNA synthesis also initiates accurately at both an early and an intermediate adenovirus promoter site.
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Cited by
Melton, D. A., Krieg, P. A., Rebagliati, M. R., Maniatis, T., Zinn, K., & Green, M. R. (1984) · Nucleic Acids Research · doi:10.1093/nar/12.18.7035 · Cited by 6,324
Mechanism of activation of a human oncogene
Tabin, C. J., Bradley, S. M., Bargmann, C. I., Weinberg, R. A., Papageorge, A. G., Scolnick, E. M., Dhar, R., Lowy, D. R., & Chang, E. H. (1982) · Nature · doi:10.1038/300143a0 · Cited by 1,393
In vivo sequence requirements of the SV40 early promoter region
Benoist, C., & Chambon, P. (1981) · Nature · doi:10.1038/290304a0 · Cited by 1,074
Klungland, A. (1997) · The EMBO Journal · doi:10.1093/emboj/16.11.3341 · Cited by 788
Two distinct transcription factors bind to the HSV thymidine kinase promoter in vitro
Jones, K. A., Yamamoto, K. R., & Tjian, R. (1985) · Cell · doi:10.1016/0092-8674(85)90113-8 · Cited by 759
Metadata
- Authors
- Manley, J. L., Fire, A., Cano, A., Sharp, P. A., & Gefter, M. L.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1980-07
- Volume
- 77
- Issue
- 7
- Pages
- 3855-3859
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W1970977612
- Cited by
- 1,211
- References
- 0
- FWCI
- 43.39
- Type
- journal article
- Language
- en
- Open access
- green
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- CoChat. (2026). Citation check: DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract. https://app.cochat.ai/papers/10.1073/pnas.77.7.3855
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- Manley, J. L., Fire, A., Cano, A., Sharp, P. A., & Gefter, M. L. (1980). DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.77.7.3855
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