Donkey made his debut in the 2001 film Shrek at a sale of mythical characters from old fairy tales, being sold to the evil Lord Farquaad's knights. Donkey's special power was his ability to talk, which his owner, an old woman, tried to capitalize on. Following a lucky escape, he befriends Shrek, who, although annoyed by Donkey's non-stop chatter, slowly and reluctantly develops an affection for him.
Donkey requests to stay with Shrek since he has no one else. At first, Shrek refuses, before finally giving in, although he only allows him to stay outside of his house for one night. Donkey denies any involvement when Shrek finds all the creatures from fairy tales living in his swamp. The creatures admit that Lord Farquaad drove them out of that location. Donkey is the only one who is prepared to lead him, and since he knows where Farquaad is, Shrek decides to accompany him.
However, when they arrive, Farquaad has his men try to kill Shrek in an arena to find the perfect champion to find and bring him back Princess Fiona, whom he wants to make his bride. Donkey and Shrek defeat Farquaad's men, and Farquaad decides to have Shrek be his champion instead. However, Shrek shows no interest and only demands his swamp back. When Farquaad offers Shrek a deal to return him his swamp if he agrees to go on the quest, Shrek agrees, and Donkey follows Shrek on his quest to find Fiona. While coming to the castle, Donkey unintentionally catches the unwanted attention of the fire-breathing dragon guarding the Princess.
Shrek escapes, but Donkey is caught and cornered. Before the dragon attempts to doom him, he tries to avoid his fate by complimenting the dragon, whom he discovers is a female, and the dragon is greatly flattered by Donkey's compliments. She takes him to her lair to be intimate with him. However, Shrek rescues Donkey before things become too personal, and the two narrowly escape with the Princess before the dragon catches up.
One night, during camp, Donkey asks Shrek why he hates everyone so much, and Shrek angrily reveals that everyone judges him as a scary monster before getting to know him. Donkey acknowledges that he already knew that there was more to Shrek's character when they met. Donkey begins to notice a romance between Fiona and Shrek, despite their denials. The next night, Donkey goes inside a windmill to talk to the Princess, only to discover that she becomes an ogre herself after sunset. Shrek overhears them talking and misinterprets the conversation as Fiona insulting him. The next day, Shrek berates her for what she supposedly said, and Lord Farquaad comes to take her.