Cloning and expression of a new Tibetan hulless barley (Hordeum vulgare) beta-1,3-glucanase gene.
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.
Yu
Zeng
Z
College of Life Science, Sichuan University, Chengdu 610064, China.
Ying
Xu
X
Lin
Tang
T
Xiao-Lin
Qiang
Q
Fang
Chen
C
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D020536:Enzyme Activators; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D043326:Glucan 1,3-beta-Glucosidase; D001467:Hordeum / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000145:classification / Q000235:genetics; D012639:Seeds / Q000201:enzymology / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D001617:beta-Glucosidase / Q000096:biosynthesis* / Q000737:chemistry* / Q000145:classification / Q000235:genetics
D020536:Enzyme Activators; D011994:Recombinant Proteins; D001617:beta-Glucosidase; D043326:Glucan 1,3-beta-Glucosidase