家蚕幼虫期和蛹期马氏管差异蛋白质组学分析  

Proteomic Comparison on Malpighian Tubules Between Larvae Stage and Pupal Stage of Silkworm(Bombyx mori)

作　　者：邹勇[1] 钟晓武[1] 张利平[1] 赵萍[1] 夏庆友[1] 

机构地区：[1]西南大学蚕学与系统生物学研究所/家蚕基因组学国家重点实验室,重庆400716

出　　处：《中国农业科学》2012年第9期1833-1839,共7页Scientia Agricultura Sinica

基　　金：国家基础研究计划(2012CB114600);国家自然科学基金项目(30972147);西南大学研究生科技创新基金优博项目(Kb2009003)

摘　　要：【目的】深入了解家蚕幼虫期与蛹期马氏管蛋白质组的变化,为马氏管在不同发育时期的功能研究提供蛋白质水平上的依据。【方法】利用蛋白质双向电泳技术对家蚕5龄第5天及化蛹第1天的马氏管组织进行比较,并采用基质辅助激光解析飞行时间质谱(MALDI-TOF-MS)对表达量差异较大的蛋白点进行肽指纹图谱鉴定,应用GPMAW 8.00软件对NCBI蛋白质数据库中所有来自于P50/Dazao的蛋白质与家蚕9倍基因组预测蛋白质库共同构建本地数据库,对试验采集到的肽指纹图谱进行分析。【结果】共检测到360—430个蛋白点,主要分布在分子质量15—80 kD、等电点4.0—9.5。2个时期共成功鉴定17个表达量有显著差异的蛋白质,其中包括与能量代谢相关蛋白质、热激蛋白、V型H+ATP合酶、30K蛋白以及与肾脏功能密切相关的丙氨酸-乙醛酸氨基转移酶2,3-羟基异丁酸脱氢酶等。【结论】本研究中差异蛋白质的发现和鉴定为完全变态昆虫幼虫期与蛹期马氏管的功能差异提供了蛋白质水平上的理论依据。[ Objective ] The objective of this study is to get more information of malpighian tubules on the proteome changes between larvae stage and pupal stage, and to provide new evidences on the protein level at different developmental stages of malpighian tubules of B. mori. [ Method ] The matrix-assisted laser desorption ionization time of flight mass spectrometry were applied for identifying the different spots which had more expression of the 2D map, and all the protein sequences from P50/Dazao on NCBI combined silkworm proteins database were used to build local-database by the software GPMAW 8.00. GPMAW was also used to analyze peptide fingerprint masses. [Result] About 360-430 spots were obtained by silver staining from the malpighian tubules after 2D-PAGE. Most of them were distributed in the area from 15 to 80 kD with pI 4.0-9.5. Seventeen markedly different proteins between larvae stage and pupal stage were successfully identified, including not only the proteins in association with energy metabolism, but also heat shock proteins, vacuolar-type W-transporting ATPase, 30K lipoprotein, and alanine-glyoxylate aminotransferase 2,3-hydroxyisobutyrate dehydrogenase which have important functions in mammal kidney. [Conclusion] The discovery and identification of these proteins can offer a valuable insights into functional differences of larvae stage and pupal stage of holometabolic insects.

关 键 词：家蚕 马氏管 双向电泳 MALDI—TOF—MS 

分 类 号：S881.2[农业科学—特种经济动物饲养]