Jacqueline Pearce starred in two Hammer horror films, The Plague of the Zombies and The Reptile, which were filmed simultaneously on the same location and both released in 1966. Other film roles include Sky West and Crooked (1965), the Carry On film Don't Lose Your Head (1966), Don't Raise the Bridge, Lower the River (1968), White Mischief (1987), How to Get Ahead in Advertising (1989), and Princess Caraboo (1994).

Despite having appeared in dozens of films and TV shows, she remains best known for her role as Servalan in the British science fiction TV series Blake's 7 (1978–1981).

As well as appearing in the BBC children's programmes Dark Season (alongside Kate Winslet) and Moondial, Jacqueline Pearce has appeared in the Doctor Who serial The Two Doctors as Chessene, a bloodthirsty alien. She has more recently been associated with Doctor Who through her appearance in The Fearmonger as Sherilyn Harper, an audio drama by Big Finish Productions, and as Admiral Mettna in the webcast story Death Comes to Time. Pearce returned once again to Doctor Who from 2015, this time opposite John Hurt, as a regular in the Big Finish audio series based on the adventures of the War Doctor. She made a memorable appearance playing the role of Rosa Dartle in the 1974 BBC series of Charles Dickens's David Copperfield. She also appeared as an associate of the Assassin Carlos in the 1988 television movie, The Bourne Identity.