Abstract
This chapter discusses the in vitro synthesis of virus-specific RNA using transcriptional complex isolated from adenovirus-infected cells. The control mechanisms of regulation of transcription and post-transcriptional modifications of RNA in eukaryotes are largely undefined. As an approach to understanding these mechanisms, a cell-free system containing an active viral-transcription complex from adenovirus-infected HeLa cells was developed. The complex contains viral DNA, cellular RNA polymerase II which has initiated transcription, methylase(s) and probably a poly (A) polymerase, and some other enzymes of processing. It appears that by proper manipulations, such a system could be used to transcribe and process RNA to generate translatable mRNA. In the presence of four nucleoside triphosphates, the complex synthesizes adenovirus-specific RNA in high yield. Depending on the experimental conditions, RNA synthesis could be carried out by elongation of already initiated RNA chains present in the complex or by de novo initiation and elongation of RNA chains in the presence of exogenous viral DNA.
Published Version
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