In vitro transcription of c-jun gene using fractionated nuclear extract from regenerating rat liver.

Regenerating rat liver serves as a source of proliferating cells, such a system can be used to study the regulation of genes involved in proliferation. We have established an in vitro transcription capable of supporting accurate transcription of cloned c-jun gene using fractionated nuclear extract prepared from partially hepatectomized rat liver. EcoR I linearized plasmid -132/+170 jun-CAT containing c-jun gene promoter region (-132 to +170 nucleotides) was transcribed in an in vitro run-off transcription assay and a transcript of expected size i.e. 370 nucleotides was obtained. The intensity of the transcript was dependent on the concentration of fractionated nuclear extract as well as template. The transcription was α-amanitin sensitive indicating that it is directed by RNA polymerase II.