DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract.
AUTOR(ES)
Manley, J L
RESUMO
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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=349725Documentos Relacionados
- Chromatin assembly in a yeast whole-cell extract
- Transcriptional activation in an improved whole-cell extract from Saccharomyces cerevisiae.
- Specific initiation by RNA polymerase I in a whole-cell extract from yeast.
- Identification of Leuconostoc spp. by analysis of soluble whole-cell protein patterns.
- Role of DNA-Dependent RNA Polymerases II and III in Transcription of the Adenovirus Genome Late in Productive Infection