关键词: 葡萄, 果实, β–葡萄糖苷酶1, 脱落酸, 基因, 克隆, 原核表达

Abstract: To explore the activity of β-glucosidase（VvBG1）in grape berries，in the present study，VvBG1 gene was first cloned through reverse transcription technique from mesocarp during Hamburg grape berry red-coloring stages. The 1 518 bp sequences encoded a polypeptide with 505 amino acids. Bioinformatics analysis showed that VvBG1 contains both a putative transmembrane region and a glycosyl hydrolase1 superfamily domain. The coding sequence after remove of its signal coding region was cloned into prokaryotic expression vector pET28a（+）and then transformed into E. coli strain BL21（DE3）by restriction endonucleases BamHⅠand XhoⅠ. The recombinant strains were successfully induced to express fusion protein and permitted to purify the VvBG1 protein. A 0.8 mg · mL^-1 of soluble protein solution was obtained after dialysis and ultrafiltration. The results gained from a combination of purified protein incubation experiment using grape berry pulp and abscisic acid（ABA）content analysis by GC-MS demonstrated that the grape berry β-glucosidase VvBG1 is of high physiological activity.

Key words: grape, berry, β-glucosidase 1, ABA, gene, cloning, procaryotic expression