论文部分内容阅读
为了进行葡萄种子蛋白质组学的研究,以葡萄种子为试材,通过对比粗蛋白质不同溶解方法、IPG胶条、上样量、等电聚焦参数以及不同SDS-PAGE分离胶浓度对双向电泳效果的影响,建立了适合葡萄种子的双向电泳体系。结果表明:TCA-丙酮提取蛋白质经酚抽提后蛋白质溶解快、杂质少、浓度高,筛选获得17cm pH 5~8的IPG胶条,上样量600μg,第一向的聚焦步骤达到80 000Vh以及11%的SDS-PAGE分离胶的双向电泳参数,蛋白质分离效果好。所建立的双向电泳体系,适用于不同葡萄品种的种子蛋白质分离。
In order to study the grape seed proteomics, grape seeds were used as experimental materials. By comparing different solubilization methods of crude protein, IPG strips, loading amount, isoelectric focusing parameters and different SDS-PAGE separation gel concentrations, Influence, established a two-dimensional electrophoresis system suitable for grape seeds. The results showed that the protein extracted by TCA-acetone had the advantages of fast dissolving protein, low impurity and high concentration. The 17g pH 5 ~ 8 IPG strips were screened, the amount of sample was 600μg, the first focusing step was 80 000Vh, 11% SDS-PAGE gel separation two-dimensional electrophoresis parameters, protein separation is good. The established two-dimensional electrophoresis system, suitable for different grape seed protein separation.