The invention belongs to the technical field of plant genetic engineering, and particularly relates to application of soybean gene promoters pEIF1 and pEIF1-I in soybean, arabidopsis thaliana and tobacco. According to the invention, a gene RPS28 and a gene EIF1 are cloned from soybeans; a promoter pEIF1 and a promoter pEIF1-I are obtained through a CTAB method, then a soybean stable transformation vector is constructed, expression of pEIF1 and pEIF1-I driven GUS protein in all stages of soybean genetic transformation is achieved, it is found that pEIF1 and pEIF1-I driven GUS is expressed in cotyledons, radicles, germs, true leaves, compound leaves, buds, petioles, internodes, roots and root nodules, it is indicated that pEIF1 and pEIF1-I are pan-expression promoters, and the activity of the expression product of the GUS protein in the soybean is measured. In addition, the transformation of pEIF1 and pEIF1-I driven GUS protein in transformation of arabidopsis thaliana and tobacco is also realized.