通过高精度的双向电泳技术对家蚕5龄幼虫的中肠组织进行了研究,利用基质辅助质量飞行时间质谱(MALD I-TOF MS)对表达量较高的蛋白点进行了肽质量指纹图谱分析,并采用GPMAW软件对家蚕基因组预测的蛋白质数据库进行了本地构库,对所得到的肽质量指纹图谱进行了分析。结果发现,家蚕中肠蛋白质经过双向凝胶电泳和图像分析,银染后可以检测出600个以上的蛋白点,这些蛋白点主要集中在分子量15~80 kD区域,等电点3~8.5之间。MALD I-TOF MS鉴定的36个蛋白点中都有较强的肽质量指纹信号峰,其中32个蛋白点得到了成功鉴定,包括原肌球蛋白以及大量的离子转运蛋白、与代谢相关的酶、与信号传导和细胞凋亡相关的蛋白等。这一结果为进一步认识家蚕中肠组织的生理功能提供了基础信息。
High resolution two-dimensional polyacrylamide gel electrophoresis, followed by computer-assisted analysis, was applied to investigate the midgut of silkworm, Bombyx mori. The matrix-assisted laser desorption ionization time of flight mass spectrometry were applied for identifying the major spots of the 2D map. Peptide mass fingerprinting were analyzed in local database by software of GPMAW. The results showed that over 600 spots were obtained by silver stain. Most of them were distributed in the area from 15 kD to 80 kD with pl 3 -8.5. All of 36 spots excised from gel had high intensity when analyzed by MALDI-TOF MS. Among them, 32 proteins, which were involved in tropomyosin, ion transportation, metabolism, signal transduction,apoptosis et al, were identified sucessfully. The result will provide a useful tool for understand the role of midgut in silkworm.