The in vivo role of the proposed poxvirus early transcription termination signal TTTTTNT was confirmed by analysis of the RNA species made by recombinant vaccinia viruses. Premature transcription termination occurred following each of two TTTTTNT sequences present naturally within the coding region of the human immunodeficiency virus type 1 envelope gene. Alteration of the TTTTTNT sequences, without changing the encoded amino acids, resulted in production of full-length early mRNAs, improved protein expression, and a more consistent immune response.