西施舌肠蛋白质双向电泳图谱的建立

Construction of Two-dimension Electrophoresis Map of the Intestinal Proteins from Coelomactra antiquata

以西施舌的肠组织为研究对象,经破碎、裂解、离心等抽提方法提取肠组织蛋白样品,利用Bradford蛋白定量试剂盒作蛋白定量后,进行一向等电聚焦电泳和SDS-聚丙烯酰胺凝胶电泳。电泳后的凝胶用考马斯亮蓝染色,利用ImageScannerⅢ扫描仪扫描凝胶,获得凝胶图像。首次得到西施舌肠蛋白质双向电泳图谱,构建了西施舌肠蛋白质组的双向电泳技术体系。图谱分析显示,所获得的肠蛋白质等电点介于pH4～7,大部分蛋白为酸性蛋白,高分子质量及高等电点蛋白含量相对较少。

The intestinal tissue of Coelomactra antiquata was used for experimental material in this study. The protein samples of intestinal tissue were extracted by crushing, cracking, centrif- ugation and other extraction methods. The Bradford protein quantitative kit was used for protein quantification. IEF electrophoresis and SDS-PAGE electrophoresis were carried out. The obtained gel was stained by coomassie brilliant blue R350, and the protein profile was scanned using ImageScanner Ⅲ to obtain the gel image. It is the first time to get the two-dimension gel electrophoresis profile and construct the two-dimension gel electrophoresis system on the intestinal proteome of C. antiquata. The map analysis showed that the obtained intestinal protein spots distributed between pH 4m7 and the majority of proteins were acidic proteins. High molecular weight or high pH proteins distributed in a relatively small area. This study has provided a foundation for the further exploration about the physiological and biochemical mechanisms on the intestinal proteome of C. antiquata.