家蚕脂肪体蛋白质组学研究

Proteomics Analysis of Fat Body from Silkworm (Bombyx mori)

摘要:

通过高精度的双向电泳技术对家蚕末龄幼虫的脂肪体组织进行了研究，采用基质辅助质量飞行时间质谱对其中一些表达量较高的蛋白点进行了鉴定，并利用GPMAW软件结合家蚕基因组预测的蛋白质数据库构建了本地的肽质量指纹图谱数据库，对所得到的肽质量指纹图谱进行了分析。研究发现，经过双向凝胶电泳及其图象分析技术，银染可以分离出722个清晰蛋白点，这些蛋白质主要集中在分子量15～90 kD区域，等电点pH 4～8之间。MALDI-TOF-MS鉴定的41个蛋白点中都有较强的肽质量指纹信号峰，其中34个蛋白点得到了成功鉴定，其中包括了大量参与代谢的酶类、不同分子量的热激蛋白、重要的血液蛋白30K，Actin3等，这一结果对人们进一步认识家蚕脂肪体提供了有利的帮助。

Abstract:

High resolution two-dimensional polyacrylamide gel electrophoresis, followed by computer-assisted analysis, was applied to investigate the fat body of silkworm, Bombyx mori. 722 spots were obtained by silver stain from 18cm，pH 3～10 gel. Most of them were distributed in the area from 15 kD to 90 kD with pH 4～8. The matrix-assisted laser desorption ionization time of flight mass spectrometry were applied for identifying the major spots of the 2D map. A total of 41 spots were excised to identify by a combination of MALDI-TOF MS after digested with trypsin. The result showed among the 34 proteins identified, plenty of them were involved in metabolism and immunity. Additionally, Heat shock proteins, 30K proteins and Actin were also detected in the fat body of silkworm. The result will provide a useful tool for understanding the role of fat body in silkworm.