Gene expression is regulated differently in prokaryotes and eukaryotes. In prokaryotes, which lack a nucleus, DNA floats freely in the cytoplasm and transcription and translation occur simultaneously. As a result, gene expression is regulated primarily at the transcriptional level. In eukaryotes, which contain a nucleus, transcription occurs in the nucleus and is separated physically from translation in the cytoplasm. Gene expression is regulated at multiple levels, including transcriptional, post-transcriptional, and translational regulation.