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目的建立以双向电泳技术结合质谱鉴定的蛋白质组学研究方法,对β3GnT8(GalT7)基因中度表达的人胃癌细胞株SGC-7901进行研究,构建差异蛋白质组学研究平台。方法使用双向电泳、图像扫描系统及图像分析软件,选择合理pH范围及长度的固相pH梯度(IPG)胶条,对样品裂解方法、电泳上样量、电泳条件、染色方法进行优化。结果使用24cm、pH4~7的IPG胶条,得到重复性及分辨率均较好的蛋白质点,对此等电点范围的蛋白质点分布有了初步了解。结论建立了以双向电泳技术结合质谱鉴定的蛋白质组学研究方法,为下一步进行SGC-7901β3GnT8(GalT7)基因敲减亚细胞群的差异蛋白质组学研究奠定了基础。
OBJECTIVE: To establish a proteomics research method identified by two-dimensional electrophoresis combined with mass spectrometry and to study the moderately expressed human gastric cancer cell line SGC-7901 with β3GnT8 (GalT7) gene and to construct a differential proteomics research platform. Methods Two-dimensional gel electrophoresis, image scanning system and image analysis software were used to optimize the method of sample cleavage, electrophoresis sample loading, electrophoresis conditions and dyeing method by selecting pH gradient and IPG strips with reasonable pH range and length. Results 24G, IP4 strips with pH4 ~ 7 were used to obtain protein spots with good reproducibility and resolution. The preliminary results of the distribution of protein spots in the isoelectric point range were obtained. Conclusion The proteomics research method established by two-dimensional electrophoresis and mass spectrometry was established, which lays the foundation for the differential proteomics study of SGC-7901β3GnT8 (GalT7) knocked-down subpopulation in the next step.