一种新型植物毒蛋白--蒜头果蛋白的基因克隆与结构初步分析

项目名称： 一种新型植物毒蛋白--蒜头果蛋白的基因克隆与结构初步分析

项目编号： No.21502166

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 环境科学、安全科学

项目作者： 袁青梅

作者单位： 云南大学

项目金额： 21万元

中文摘要： 课题组在前期研究中，从蒜头果种子中分离得到一种新的植物毒素--蒜头果蛋白（Melanin），明确了Melanin是II型核糖体失活蛋白（RIPS），由A、B两条链通过一对二硫键相连，并确定两条链N末端10个氨基酸序列。活性研究发现Melanin对HeLa细胞的IC50为1.26nmol/L，并有显著的诱导癌细胞凋亡的作用。但对于这一潜在的药用蛋白，还知之甚少，对它的一级结构也几乎一无所知。因此，本项目拟通过构建cDNA文库及PCR技术，筛选目的基因片段，利用3'-RACE和SMART-RACE获取malanin全长cDNA基因，推测氨基酸序列。在materials studio（MS）软件的Forcite等模块下来实现分子模拟，分析其合理的高级结构及性质。获知malanin翻译后加工剪切的情况。为今后构效的研究奠定基础。

中文关键词： 蒜头果蛋白；基因克隆；氨基酸序列；结构分析

英文摘要： In the previous research, Melanin, a new toxin,was isolated from seeds of Malania oleifera by our research group. And it was found that Melanin is a type II Ribosome-inactivating proteins (RIPS), composed of chain A and B that are connected by a pair of disulfide bonds. 10 amino acid has been determined at the N-Terminal of the two chains. Activity test showed that its IC50 on HeLa cells is 1.26nmol/L and has a significant role of inducing apoptosis in cancer cells. But, we still know the potential pharmaceutical protein too little, even nothing about its amino acid sequence and structure. Therefore, this project plan to filter the gene fragment of Malanin through the construction of cDNA library and PCR techniques.And also we will use 3'-RACE and SMART-RACE to obtain full length cDNA gene of Malanin. Then to speculate the amino acid sequence. Finally, the molecular simulation on MS software is used to predict the advanced structures and properties. We hope that this study can become the foundation for the research on the link between structure and activity of Malanin in the future.

英文关键词： malanin; gene cloning;The amino acid sequence;structural analysis