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为建立适用于显性多子房小麦细胞质效应的蛋白质双向电泳体系,以显性多子房小麦材料DUOII与特异细胞质材料Te Zhi I杂交的F1幼穗为材料,采用TCA-丙酮法提取蛋白质,并在IPG胶条长度和p H范围、SDS-PAGE凝胶浓度及蛋白质上样量等方面,对多子房小麦幼穗蛋白质双向电泳体系进行了探究与优化.结果表明,本文采用的蛋白质定量方法准确度高(R2=0.9999),确立了17 cm,p H4~7的IPG胶条,12%SDS-PAGE分离胶,上样量为900μg的双向电泳方法体系,获得了最适合本研究蛋白质组分析的双向电泳图谱.经PDQuest 2DE 8.0.1软件分析,2-DE图谱上可分辨出1 444±14个清晰蛋白质点,且重复性较高(95%),相关系数为0.960.建立了一套适用于显性多子房小麦细胞质效应研究的蛋白质双向电泳体系.
In order to establish a two-dimensional electrophoresis system suitable for the cytoplasmic effect of dominant multi-ovary wheat, the F1 spike hybrids of DUOII, a specific multi-ovary wheat material, and Te Zhi I, a specific cytoplasmic material, were extracted by TCA-acetone method. The two-dimensional electrophoresis system of wheat young spike protein was explored and optimized in terms of IPG strip length and p H range, SDS-PAGE gel concentration and protein loading, etc. The results showed that the protein quantification The method has high accuracy (R2 = 0.9999), IPG strips with 17 cm and p H4 ~ 7, 12% SDS-PAGE separation gel and 900 μg sample volume were established. Two-dimensional electrophoretograms were analyzed by PDQuest 2DE 8.0.1 software, 1 444 ± 14 distinct protein spots were resolved on the 2-DE map with high reproducibility (95%) and a correlation coefficient of 0.960. A Two - dimensional Protein Electrophoresis System Suitable for Study on Cytoplasmic Effect of Dominant Wheat.