Mitogen-activated protein kinase (MAPK) cascade constitutes a conserved signaling module in eukaryotes. MAPK kinase (MAPKK) plays a crucial role in a MAPK cascade. ZmMEK1 is the first characterized MAPKK gene in maize. Although ZmMEK1 has been studied in detail in biochemical level, the genomic organization of ZmMEK1 gene is obscure. In this research, we clarified ZmMEK1 is a single-copy gene in the maize genome. Southern blot analysis using 3' specific region of ZmMEK1 cDNA as a probe revealed the presence of distinct single bands in each lane of EcoRI and HindIII. Although previous Southern blot analysis using full-length ZmMEK1 cDNA as a probe revealed several hybridizing bands, we showed here that all bands come from one genomic fragment corresponding to ZmMEK1 gene. Furthermore, ZmMEK1 was induced by PEG, abscisic acid (ABA), and salicylic acid (SA) and was down-regulated by NaCl.