Four B-hordein genes, designated BH1-BH4, were cloned using PCR amplification from two hull-less barley cultivars, ZQ7239 and ZQ148, collected from Tibet. The results of sequencing indicated that BH1-BH4contained complete open reading frames (ORFs). Comparison of their predicted polypeptide sequences with the published sequences suggested that they all share the same basic protein structure. Phylogenetic analysis indicated that the deduced amino-acid sequences of BH1-BH4genes were more closely related to B-hordeins from cultivated barley ( Hordeum vulgareL.) than to any other prolamins from wild barley and Aegilops tauschii. Comparison of the coding regions of BH1-BH4genes showed that BH1had a lower sequence identity to other previously published B-hordeins than the other three B-hordeins obtained in this study. BH1was then cloned in a bacterial expression vector based on bacteriophage T7 RNA polymerase. The resulting plasmid produced a 28.15 kDa protein in Escherichia coli. The potential value of B-hordein genes in grain quality improvement of hull-less barley has been discussed.