To evaluate qualitative and quantitative expression level of HMW-GS protein, we separated glutenin fractions andconducted two-dimensional electrophoresis (2DE) in 32 Korean wheat cultivars. The average spot numbers of HMW-GS inKorean wheat cultivars were 11.78 which included 1.31, 5.53 and 4.94 spots at Glu-A1, Glu-B1 and Glu-D1 loci, respectively.Cultivars harboring 1, 2* subunits had more spots than ones with null allele in Glu-A1 locus because Glu-B1 and Glu-D1 locidid not show difference in spot number. A large variation in the total spot number of HMW-GS was observed ranging from7 (Dahong) to 18 (Jokyung). Relative expression level of Korean wheat cultivars when compared with the check accessionChinese Spring ranged from 24% (Eunpa) to 111% (Gobun) with an average of 44%. Korean wheat accessions could be classifiedinto six groups based on the number of HMW-GS spots and quantification value of each spot.