茳芒决明抗菌蛋白的分离纯化及当归和六种决明属植物的毛细管电泳指纹图谱-生物化学及分子生物学专业论文.docxVIP

茳芒决明抗菌蛋白的分离纯化及当归和六种决明属植物的毛细管电泳指纹图谱 茳芒决明抗菌蛋白的分离纯化及当归 和六种决明属植物的毛细管电泳指纹图谱 生物化学及分子生物学专业 研究生：宋贤丽 指导教师：刘克武 摘要本研究对103种药用植物材料进行了抗菌蛋白的初步筛选工作，确定其 中有17种材料可能有抗菌蛋白。对茳芒决明进行了抗菌蛋白的分离纯化：经粉 碎、磷酸缓冲液浸提、硫酸铵沉淀、CM-Sepharose FF XK 26阳离子交换层析、 两次Mono S阳离子交换层析、Superose 12HR分子筛层析可得到具抗真菌活性 的蛋白。纯化的抗菌蛋白在SDS上显示单一条带，亚基相对分子质量为 28．4kD。 利用毛细管电泳来分析了六种豆科决明属药用植物种子中的水溶性次生代 谢物。电泳分离缓冲液为0．1mol／LSDS，0．1mol／L硼酸盐(pH8．5)，运行电压为 25kV。确定4个峰为六种决明属植物种子的共有峰，并得到各自的特征峰。六 种决明属植物种予的毛细管电泳指纹图谱有明显的差异，可利用这些差异对其 进行鉴别，并为以后这些种子中次生代谢物的定性定量工作打下基础。 利用毛细管电泳来分析了八个不同产地的当归中的水溶性次生代谢物。以 岷县当归为道地药材，确定氓县当归有19个共有指纹峰；八个不同产地的当归 有9个共有指纹蜂。通过相似度的计算可知，四川南坪和宝兴，甘肃渭源和云 南丽江的当归质量和岷县当归较近，青海渭中、湖北利川和建始的当归质量和 岷县当归相差较远。 关键词茳芒决明；抗菌蛋白；分离纯化：决明属；当归：次生代谢物；毛细管 电泳 Isolation Isolation and purification of the Antifungal Protein from Cassia sophera Linn．and the Capillary Electrophoresis fingerprint ofAngelica sinensis(Oily．) and Cassia seeds of six species Major：Biochemistry and Molecular Biology Graduate：Song Xianli Instructor：Liu Kewu ● ABSTRACT We have sifted 103 medicinal plants．roughly idemified 17 plants mi曲t contain antifungal proteins．Antifungal protein was purified from Cassia sophera Linn．by extraction，fraction with(NH4)2S04，cation-exchange chromatography of CM·Sepharose FF XK 26，the first cation-exchange chromatography of Mono S and the second one，followed by gel filtration of Superose 12HR．ne purified antifungal protein gave one band on SDS—PAGE，and the subunit molecular mass is about 28．4kD． The water-soluble secondary metabolites of Cassia seeds of six species were analyzed by capillary electrophoresis．The running buffer was O．Imol／L borne． O．1mol／LSDS，pH 8．5，and the separation voltage Was 25kV．Four con3_rnon peaks Can be found in the electropherograms of Cassia seeds of six species，the charactedsfic peaks can also be observed．Fingerprint of the Cassia seeds of six species showed significant differences．which c觚be used for their identification． Then we analyzed water-soluble secondary metabolites of Angelica sinensis rof州Diels from eight different