双向电泳法在家蚕蛋白质分离中的应用  

The application of two dimensional electrophoresis in the separation of proteins in Bombyx mori

作　　者：易发平[1] 张平波[2] 常平安[3] 宋方洲[1] 李斌[4] 藤井博[2] 

机构地区：[1]重庆医科大学生物化学与分子生物学教研室 [2]日本九州大学遗传资源中心 [3]重庆邮电学院生物信息学院,重庆400065 [4]西南大学农业部蚕桑学重点实验室,重庆400716

出　　处：《昆虫知识》2006年第6期873-876,共4页Entomological Knowledge

基　　金：国家自然科学基金项目(NO.30371479);重庆市骨干教师资助计划(渝教人200415);重庆市科委项目(渝科发计字[2002]18号)

摘　　要：家蚕Bombyxmori(L.)既是重要的经济昆虫,又是鳞翅目昆虫研究的典型模式生物。开展家蚕蛋白质组研究,将有助于阐明家蚕绢丝蛋白的分泌机理,也是研究鳞翅目昆虫及其他生物生命本质的需要。双向电泳是蛋白质分离的关键技术。为探讨适宜家蚕蛋白质组研究的双向电泳条件,以家蚕丝腺、丝腺内容物、蚕卵和血液为材料,在不同条件下进行双向电泳,并对分离的蛋白点进行质谱分析。结果表明:通过改进的蛋白质裂解液辅以超声破碎制备的蛋白质,双向电泳后能够得到较好的2-DE图,也能满足进行MALDI-TOFMS分析的需要。因此本研究方法适用于家蚕不同组织中蛋白质的提取和双向电泳。The silkworm, Bombyx mori （L.）, is not only an important economic insect but also a typical animal model for research on lepidopteron insect. The study on the proteome of B. mori will help to explain the secretion mechanism of silk proteins. Moreover, the study on the proteome of B. mori is necessary for studying on the life essence of lepidopteron insect and other organisms. The two dimensional electrophoresis（2-DE） is the key skill to isolate proteins. In order to get suitable 2-DE circumstance for the proteome study, the proteins were extracted from the silk gland, internal substance of silk gland, eggs and blood, separated by 2-DE under the difference circumstances and identified by peptide mass fingerprinting method. The results indicated that a 100 p.g protein sample prepared with lysis buffer and ultrasonic cell disruptor was sufficient for 2-DE and the protein spots were separated very well for MALDI-TOF MS. Therefore, the method is suitable for extraction of the proteins of B. mori from different tissues and 2-DE.

关 键 词：家蚕 双向电泳 蛋白质组 

分 类 号：S881.2[农业科学—特种经济动物饲养]