蓖麻毒蛋白的提纯及杀虫效果研究-生化与分子生物学专业论文.docxVIP

摘要 离麻毒蛋白 Ricin 是存在于蔑麻种子中的一种毒蛋白 F 分子量在 64000，由 A，B 两条肤链基组成 p 通过二硫键相连。 A 链是活性链，分子量在 31 000，B 链分子量在 34 000，具有凝集素活性，能与半乳糖残基结合。 A 链通过 B 链特 异地结合细胞表面的受休，通过经受体介导的内吞方式进入细胞。 A 链能与核糖 体的 60S 大亚基结合贯通过其特异的 N瞩糖音酶活性，催化切断 60S 亚基 28S rRNA 中第 4234 位腺嘿岭与核糖分子之间的糖昔键，这一喋岭的脱落可导致整个核糖 体的失活?从而影响需核糖体参与的氨基胁 tRNA 的结合和肤酶t刚A 的转位过 程，最终导致蛋白质合成障碍 F 引起细胞的死亡。蔑麻毒素具有强烈的细胞毒性， 因此具有抗肿瘤和开发生物农药的前景。 本文研究了从蔑麻种子中分离纯化蔑麻毒蛋白的方法 F 对蔑麻籽粗提物和离 麻毒蛋白进行了杀虫研究。经硫酸锁盐析，亲和层析和凝胶过滤，得到了纯的蔑 麻毒蛋臼量在 sos幡PAGE 中F 显示出分子量在 64 000 左右，存在两个亚基 p 分 子量分别为 31 000 和 34 000，与文献报道相符合。用蔑麻籽粗提物和毒蛋白对 妨虫和菜青虫进行毒力测定费结果表明 p 黯麻毒蛋臼具有一定的触杀毒性声对菜 青虫胃毒活性弱声离麻碱具有较强的胃毒，触杀活性，并对菜青虫具有较高的拒 食活性。 关键词:蔑麻毒蛋白 分离纯化菜青虫毒性  Abstract The ricin is a plant toxin that can be iso 1ated from castor beans (Ricinus communìs). The protein toxin has a molecular weight of about 64 000，which consìsts of two chains linked by a disulfide bond. Ricin A chain (RTA) is a ribosome-inactivating protein，whose molecular weight is about 31 000. Rìcìn B chaio is the binding subunit and can specifically recognize receptors on the cell rnembrane ，which have a terminal galactose. RTB molecular weight is about 34 000. The ricin enters into the cell through receptor-mediated endocytosis. RTA binds to protein L3 of ribosomal 60S subunit. It has specific N-glycosidase activity，and can catalytically cleave a specific adenine at the site of 4 234 out of the 28S rRNA. This cleavage of adenine can lead to the disruption of ribosomal funct?on，thereby，iohibits the protein synthesis，and then cause the death of cells. For this reason，ricin has a good future 岛r applying on anti-tumor and bi010gical pesticide. Experimental methods and technology for the separation and purification of ricin from castor beans were described in this paper. By ammonium sulfate precipitation ， Sepharose-4B affinity chromatography ，and followed by molecular sieve chromato -graphy ，purified rìcin was obtaìned. The purified preparation showed two protein bands with relative molecular weights of31 000 and 34 000. Eff