Prolonged Transcription in a Cell-Free System Involving Nuclei and Cytoplasm

AUTOR(ES)

Wu, Guang-Jer

RESUMO

A cell-free system for nuclear-directed transcription has been developed that gives prolonged synthesis in the presence of cytoplasm. The nuclear and cytoplasmic components have been prepared from Krebs II ascites tumor cells for most experiments but further observations indicate that components prepared from other cell types may be used. After an initial 5- to 10-min period of relatively rapid RNA synthesis a linear rate ensues for 2-3 hr. In the absence of cytoplasm no net RNA synthesis occurs after the initial 10-min period. Experiments with α-amanitin suggest that about half of the cell-free synthesized RNA is made by RNA polymerase II, the enzyme believed to be responsible for messenger synthesis in vivo.

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