Transcriptionally active nuclear extracts have been prepared from rat liver, brain, and spleen. The adenovirus-2 major late promoter directs efficient transcription by RNA polymerase II in all of these extracts, whereas the promoter of the mouse albumin gene is significantly used only in the liver extract. Albumin sequences located between -170 and -55 are required for this liver-specific in vitro transcription, since deletion of this region results in almost a 100-fold reduction in transcription. In addition, insertion of these sequences in either orientation upstream of the parotid-specific Amy-1 promoter, which is poorly transcribed in the liver extract, increases the activity of this promoter to a level comparable to that observed for the albumin promoter.