Abstract:WRKY is a class of transcription factors related to plant stress-tolerance, which improve plant resistance by activating expression of a series of stress response gene downstream. In this study, we screened a stress-induced WRKY transcription factor gene GhWRKY2 from Gossypium hirsutum based on the gene chip, it belonged to WRKYⅡ subgroup. We obtained the full length of GhWRKY2 cDNA by RT-PCR technology, and made sequence analysis and structure-function prediction by bioinformatics software. According to the sequence homology of GhWRKY2 protein, a multiple sequence alignment and phylogenetic tree construction of homologous species was made by MEGA7. Amino acid the physical and chemical properties was analysised by ProtParam and Motif scan; protein sequence was analysised by SMART online tools; the structure and tertiary structure of GhWRKY2 gene were analyzed by SWISS-MODEL online tool; the structure and tertiary structure of GhWRKY2 gene were analyzed by SWISS-MODEL online tool. The results showed that the GhWRKY2 contained a 942 bp open reading frame encoding a 313 amino acids residues with a predicted molecular mass of 34.22 kD and an isoelectric point of 8.76. The transient expression of GhWAKY2-GFP into Arabidopsis protoplasts showed that the GhWRKY2 was located in the nucleus. Moreover, it was found to have transcriptional activation activity. The clone of stress response GhWRKY2 gene, lay the foundation for further confirming the biological function of resistance at the molecular level, and revealing mechanism of abiotic stress resistance in cotton, and providing new candidate genes for stress tolerance breeding in cotton.