DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract.

Abstract

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.

Keywords

Transcription (linguistics)BiologyMolecular biologyDNARNARNase PRNA polymerase IIRNase HPromoterGeneGene expressionBiochemistry

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Publication Info

Year: 1980
Type: article
Volume: 77
Issue: 7
Pages: 3855-3859
Citations: 1282
Access: Closed

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APA Style

                            
                                
                                    James L. Manley, 
                                
                                    Andrew Fire, 
                                
                                    Amparo Cano
                                
                                et al.
                            
                            (1980). 
                            DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract.. 
                            Proceedings of the National Academy of Sciences
                            , 77
                            (7)
                            , 3855-3859.
                            https://doi.org/10.1073/pnas.77.7.3855
                        

Identifiers

DOI: 10.1073/pnas.77.7.3855