[The function of T7 promoter as cis-acting elements for polymerase II in eukaryotic cell]

Using the chlorampheniol acetyltransterase gene as reporter, the function of phage T7 promoter in mammalian cells was studied by inhibition of transcription with alpha-amanitin. The experiment proved that the reporter under T7 promoter was transcribed by RNA polymerase II. Competitive electropho retic mobility shift assay (CEMSA) with TATA box, CAAT box, GC box and octamer showed that the TATA box was competitive molecular for synthetic T7 promoter. It is possible that T7 promoter is bound with TF II D transcription factor. The TATA box and octamer were inserted into Pvu II site upstream from the T7 promoter of pT7CAT. Two recombinant plasmids, pT7TATACAT and pT7OCTCAT, were constructed and transfected into CHO cells. CAT-activity test showed that T7 promoter strength was increased by octamer factor, not by TATA box. These results suggested that T7 promoter functions as cis-acting elements of RNA polymerase II transcriptional system in eucaryotic cells.