H. Kajiwara, A. Imamaki, Masatoshi Nakamura, K. Mita, Q. Xia, M. Ishizaka
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引用次数: 7
摘要
对家蚕(Bombyx mori L.)第5龄第3天幼虫进行了脂肪体蛋白质组学分析,并从第4龄第1天幼虫到成蛾前,用二维聚丙烯酰胺凝胶分离蛋白的表达情况进行了跟踪。手工切除2d凝胶上的蛋白质并用4-乙烯基吡啶进行烷基化处理。每个斑点经胰蛋白酶水解后,采用毛细管高效液相色谱-离子阱质谱联用分析。在之前的报道1)中,我们使用果蝇基因组中获得的氨基酸序列数据库进行蛋白质鉴定。利用家蚕基因组的氨基酸序列数据进行分析。此外,将脂肪体中的蛋白质表达与蚕蛋白质组数据库(http://kaiko2ddb.dna.affrc.go.jp/cgi-bin/search_2DDB.cgi)中描述的其他阶段和其他组织中的蛋白质表达进行了比较。从先前的报道中鉴定出几个未确定的蛋白质(1)具有较高的Mascot分数。在变形过程中,蛋白质的诱导和减少被观察到,在网络上连续的2d凝胶上,斑点浓度的变化。
Proteome analysis of silkworm (Bombyx mori L.) fat bodies was performed on the fifth-instar day-3 larva stage and the expression of proteins separated on two-dimensional polyacrylamide gels was tracked from the fourth-instar day-1 larva stage to the stage before adult moth. Proteins on the 2D-gels were excised manually and treated with 4-vinylpyridine for alkylation. Each spot was analyzed by capillary high-performance liquid chromatography coupled with ion-trap mass spectrometry after proteolysis using trypsin. In the previous report1), the amino acid sequence database obtained from the Drosophila genome was used for the protein identification. This analysis used the amino acid sequence data elucidated from the silkworm genome. Additionally, protein expression in fat bodies was compared with that at other stages and in other tissues described in the silkworm proteome database (http://kaiko2ddb.dna.affrc.go.jp/cgi-bin/search_2DDB.cgi). Several unidentified proteins from the previous report1) were identified with high Mascot scores. Induction and reduction of proteins during metamorphosis were observed as changes in spot concentration on sequential 2D-gels on the web.