A high titer retroviral vector containing the cDNA of human estrogen receptor (hER) was generated and used to transfer the hER gene into the rat 208F cell line. Southern blot analysis showed the integration of the provirus to be at a unique site and that the provirus was intact in the genome of recipient cells. The expression of the integrated hER gene in the infected rat cells was detected by Northern blot analysis and by a functional assay in which the hER gene product stimulated the production of a chloramphenicol acetyl transferase gene under the control of an estrogen-responsive element. These experiments demonstrate the feasibility of using a retroviral vector system to introduce a functional ER gene into cultured cells lacking this receptor.