In this paper, the NtWRKY8 gene was cloned from Nicotiana tabacum L. based on the database resources and RT–PCR technology. Sequence analysis showed that NtWRKY8 gene contained a complete ORF, which was 1 551 bp and encoded a protein of 516 amino acid residues. The WRKY8 protein had two conserved WRKY domains, and the zinc-finger structure was C2H2 (C–X4–5–C–X22–23–H–X1–H). Functional analysis indicated that this protein was probably located in the nucleus and had functions in replication, transcription and regulatory, and consequently could regulate the growth, development and response to stress in plants. The RT–PCR experiment showed that the relative expression of NtWRKY8 gene was induced under low-temperature treatment.