柴胡及体细胞杂种愈伤组织酯酶同工酶技术的优化

Optimizing of Esterase Isoenzyme Analysis of Bupleurum scorzonerifolium and Its Somatic Hybrid Calli

为解决柴胡及其体细胞杂种愈伤组织中次生代谢物多,同工酶粗提液粘稠、杂质多且呈混浊态,同工酶电泳谱带不清晰及易拖尾的问题,通过增加离心次数、调整酶粗提液上样体积等方法,得到了理想的酯酶同工酶电泳谱带。该法广泛适用于其它植物的同工酶聚丙烯酰胺凝胶电泳分析。

Numerous secondary metabolites in Bupleurum scorzonerifolium somatic hybrid calli usually cause a sticky and murky crude extract of isoenzymes,which contains many unknown impurities.Troubles with poor definition and serious smear are typically found due to an unacceptable purity of the sample.To solve the problem,multi-rounds centrifugation were undertaken and then the loading volume of the sample was adjusted to proper range.An ideal result was achieved though the optimized assay,which can be widely applied into PAGE analysis of isoenzymes in other plants.