Accurate transcription initiation in an Ehrlich ascites tumor cell lysate

An in vitro system for accurate transcription initiation by RNA polymerase II was developed using Ehrlich ascites tumor cells. Truncated DNA containing adenovirus 2 major late promoter was faithfully transcribed in this lysate, although the efficiency of transcription was lower than that in a HeLa cell lysate. Creatinephosphate greatly enhanced accurate transcription in this lysate. The transcriptions of various truncated mouse genes containing promoter regions in this lysate were tested, but the syntheses of run-off products were not clearly detected. Adenovirus 2 major late promoter was utilized more efficiently when integrated into circular plasmid DNA than when integrated into truncated DNA, as shown by S1 nuclease analysis.