A new full-length beta-1,3-glucanase cDNA was obtained by RT-PCR and RACE techniques from Tibet hulless barley and its complete gene sequence obtained by DNA Walking. Sequence alignment with the BLAST program showed that cDNA has high similarity with barley beta-1,3-glucanase II. The gene was functionally expressed in E. coli and the recombinant protein catalysed the hydrolysis of beta-1,3-glucan with an action pattern characteristic of a beta-1,3-glucan endohydrolase (EC 3.2.1.39). Southern blot analysis indicated that the gene is a member of a small gene family. RT-PCR and northern blot analysis indicated it is constitutively expressed in barley shoots.