人精子膜蛋白的二维电泳实验研究

Experimental Research on Human Spermatozoa Membrane Proteins by Two-Dimensional Gel Electrophoresis

目的 :利用二维电泳技术对人精子膜蛋白进行分离 ,为建立正常人精子膜蛋白的蛋白质图谱打下基础。 方法 :使用Percoll密度梯度离心筛选精子 ,用等电聚焦结合聚丙烯酰胺凝胶电泳对人精子膜蛋白进行二维电泳分析。 结果 :图像分析软件共可识别出约 80 0个蛋白质斑点 ,绝大多数蛋白的相对分子质量为 2 0 0 0 0～ 10 0 0 0 0 ,等电点为 3.0～ 7.0。 结论 :精子膜上至少有 80 0种蛋白 。

Objectives: To analyze human spermatozoa membrane proteins by two dimensional gel electrophoresis and to provide a basis for drawing the protein map of normal human spermatozoa membrane proteins. Methods: Spermatozoa were purified by Percoll density centrifugation, and spermatozoa membrane proteins were analyzed by two dimensional gel electrophoresis using isoelectric focusing and polyacrylamide gel electrophoresis. Results: About 800 protein spots could be identified by the imaging analysis system. The molecular weight and isoelectric point of most proteins were 20 000 to 100 000 and 3.0 to 7.0 respectively. Conclusions: There are more than 800 types of proteins in the human spermatozoa membrane. The spermatozoa membrane protein can be identified with a certain precision by two dimensional gel electrophoresis.