Transcriptional initiation of T7 promoter by transcriptional system in eukaryotic cells

Using the chlorampheniol acetyltransterase gene and the cDNA of low-density lipoprotein receptor gene as reporters, the function of phage T7 promoter in mammalian cell was studied by inhibition of transcription with α-amanitin, by transcription assay with T7 polymerase in vitro and electrophoretic mobility shift assay(EMSA). The results demonstrated that CAT and LDLR genes as reporters under T7 promoter were transcribed by RNA polymerase Ⅱ. Electrophoretic mobility shift assay revealed that the binding of synthetic T7 promoter with nuclear protein, suggesting further the function of T7 promoter as cis-acting elements of transcriptional system in eucaryotic cells. [