论文部分内容阅读
目的建立针对钉螺肝脏蛋白质组阵列分离的双向电泳方法。方法分析裂解液、细胞破碎、沉淀方法对钉螺肝脏蛋白质提取的影响,并优化双向电泳参数,建立钉螺肝脏蛋白质的提取方法和双向电泳条件。结果LysisⅢ(8mol/L尿素,4%CHAPS,65 mmol/L DTT,40 mmol/L Tris,0.5%IPG Buffer)提取钉螺肝脏总蛋白,2-D Clean-up试剂盒处理纯化总蛋白,等电聚焦参数为8000 v,50000 vh,成功获得了分辨率高、重复性好的双向电泳图谱,凝胶经EMBL银染,可分辨蛋白质斑点数约430个。结论建立了钉螺肝脏蛋白质双向电泳技术,为开展钉螺蛋白质组学研究奠定了良好基础。
OBJECTIVE To establish a two-dimensional electrophoresis method for the separation of hepatic proteomes in the liver. Methods The effects of lysate, cell disruption and precipitation methods on the protein extraction of liver were studied. Two - dimensional electrophoresis parameters were optimized to establish the extraction method and two - dimensional electrophoresis of liver protein. Results The liver total protein was extracted with Lysis Ⅲ (8 mol / L urea, 4% CHAPS, 65 mmol / L DTT, 40 mmol / L Tris and 0.5% IPG Buffer). Total purified protein was purified with 2-D Clean- With the focusing parameters of 8000 v and 50000 vh, two-dimensional gel electrophoresis with high resolution and good reproducibility was successfully obtained. The gel was identified by EMBL silver staining and the protein spots were about 430. Conclusion The two-dimensional electrophoresis technique of liver protein was established, which laid a good foundation for the study on proteomics of snail.