In this research, two EREBP (ethylene response element binding protein) genes were isolated by the yeast one-hybrid system, named GhEREB2 and GhEREB3, and both have one intron in their coding regions. The deduced amino acid sequences of GhEREB2 and GhEREB3 have some typical features of transcription factors, one potential basic nuclear-localization signal, one possible acidic activation domain, and one conserved DNA binding domain, and they show high similarity, especially in the DNA-binding domain. GhEREB2 was expressed in roots, stems, and leaves (low levels), but GhEREB3 was only expressed in leaves. Furthermore, the expression of GhEREB2 and GhEREB3 was induced by ethylene and jasmonic acid. In addition, GhEREB2 and GhEREB3 proteins specifically bind to a GCC-box and strongly activate the expression of HIS3 and LacZ reporter genes in yeast. Collectively, these results suggest that GhEREB2 and GhEREB3 might be involved as positive transcription factors in biotic stress signal transduction pathways.