带斑狼蛛粗毒的生理生化分析毕业论文.doc

带斑狼蛛粗毒的生理生化分析 摘 要 本文描述的带斑狼蛛分布于我国西南地区，常见于稻田及湿地地带。带斑狼蛛毒液是一种化学混合物，对该蜘蛛粗毒进行生物化学和生物学活性方面的分析，探究带斑狼蛛粗毒潜在的药用价值。主要实验步骤为：反相高效液相向色谱（RP-HPLC）分离得到各个组分；基质辅助激光解析电离飞行时间（MALDI-TOF）质谱分析，测定高效液相色谱分离峰的分子量；聚丙烯酰氨凝胶电泳 （SDS）技术分析粗毒中的小分子量多肽与大分子量蛋白质；粗毒的细胞毒活性分析，带斑狼蛛粗毒对白血病K562细胞、前列腺癌PC-3细胞具有显著的细胞毒活性；抗菌实验结果显示，该粗毒对金黄色葡萄球菌、绿脓杆菌、变形杆菌无抑制作用，但对谷氨酸棒状杆菌具有一定的抑制活性。 关键词 带斑狼蛛 粗毒 高效液相色谱 飞行时间质谱 聚丙烯酰氨凝胶电泳 细胞毒活性 抗菌活性 Physiological and Biochemical study of the venom of spider Lycosa vittata The wolf spider Lycosa vittata described in this article distributes in Southwest China, found in rice fields and wetland areas. The wolf spider Lycosa vittata venom is a chemical mixture, to explore whether crude venom of the wolf spider Lycosa vittata has medicinal value ,we study the biochemistry and biological activity of the crude venom, Main experimental steps as follows: high performance liquid chromatography (RP-HPLC) separates all components; matrix assisted laser analysis ionization flight time (MALDI-TOF) mass spectrum analysis determines the molecular volume of the separation peak; polypropylene ammonia coagulation rubber electrophoresis (SDS) technology analysis the small molecular volume peptides and macromolecules proteins; Cytotoxic activity of the crude venom, analysis the venom on for leukemia K562 cells and prostate cancer PC-3 cell，antimicrobial activity against for Pseudomonas aeruginosa, staphylococcus aureus ,Proteus and Glutamine Corynebacterium: results are as follows: The wolf spider Lycosa vittata is rich in peptide component and has obvious cell growth inhibition activity for leukemia K562 cells and prostate cancer PC-3 cell , The wolf spider Lycosa vittata has inhibition on Glutamine Corynebacterium. Key words: spider; Lycosatuttatasp.nov; crude venom; HPLC; MALDI-TOF; SDS; cell growth inhibition ；antimicrobial activity ; 1 绪论 1.1研究背景 蜘蛛属于节肢动物门，蛛形纲，蜘蛛目，蜘蛛起源早、种类繁多，分布广泛。全世界有近40,000种，我国大约有4, 000种，如间斑寇蛛（俗称黑寡妇蜘蛛）、虎纹捕鸟蛛、海南捕鸟蛛、中华狼蛛、雷氏大疣蛛和漏斗网蛛等，在我国剧毒蜘蛛也有10多种[1]。蜘蛛通常以昆虫与哺乳动物为食，以蜘蛛毒液来麻痹和杀死猎物，同时蜘