Cloning Of Dioscin-glycosidase Gene

In this paper, purification and gene sequence from dioscin-glycosidase were studied. For this enzyme can particularity hydrolyze rhamanopyranosyl of dioscin into glycosyl-less dioscin.Dioscin-glycosidase was successfully purified by DEAE-Cellulose DE52, molecular weight is56kDa by menstruation, and the N-terminal sequence of dioscin-glycosidase is ATPADWRSQ.The total RNA of Absidia sp.G3d was isolated. The primers were designed according to the N-terminal sequence of dioscin-glycosidase, and the unknown sequence of gene CDS was amplified by RT-PCR. Designing the primers on the basis of above sequence, the complete gene sequence was amplified by RACE. The sequence length sequenced is1725bp, and gene CDS section is1497bp.Amino acid segments of dioscin-glycosidase were sequenced by mass spectrum technology, and the segments were compared with the amino acid sequence that was from the translated gene sequence, and comparability was up to100%. So it is proved that the gene sequence above is right.