【Objective】The promoter of rice glutelin B1(pGluB1) has been intensively used to study high seed-special expression of exotic genes, and as a model to understand regulation mechanism of seed-storage protein genes. Former researchers reported that pGluB1expressed only in rice seed endosperm with no expression activity in other tissues such as root, stem, leaf, sheath and glume etc. The present study is aiming to overcome the shortcoming of time consuming for screening of seed-special expression promoters.【Methods】The expression vector pCAMBIA1302-pGluB1sig- CTBIN-NOS, in which a fusion gene of the cholera toxin B subunit and human proinsulin (CTBIN) was driven by the 2.3 kb promoter sequence of rice glutelin GluB1 with its signal peptide (pGluB1sig), was transformed into the rice calli from mature embryo via Agrobacterium-mediated method. The expression of fusion gene CTBIN at both transcription and translation level was tested by RT-PCR and Western-blotting assay.【Results】Among the seven transgenic calli clones, six clones of the fusion gene CTBIN were expressed at transcription level. The selected four clones subjected to Western-blotting assay were all fatherly verified in their expression at translation level. Additionally, according to the molecular weight, we speculated that the signal peptide (24 aa) of GluB1 at N-terminus of the fusion protein CTBIN has been excised successfully from callus cells of all the test clones.【Conclusion】The rice seed-specific expression promoter pGluB1 can drive an exotic gene expression in callus, and the seed protein body subcellular-targeted signal peptide could be excised from callus cells. The results lay a foundation of the quick detection of expression activity of plant seed-specific expression promoters in callus and exploiting the mechanism of protein subcellular sorting selection in callus cells.