摘要为建立和优化绒山羊(Capra cashmere)皮肤组织蛋白质双向电泳技术(2-DE)，本研究从样品提取及纯化方法、上样量、IPG胶条的选择、等电聚焦程序等多个方面对双向电泳分离条件进行比较研究。结果显示，液氮研磨+超声波破碎法提取皮肤组织蛋白质，2-D clean up kit试剂盒进行纯化，采用17 cm pH 4~7的IPG 胶条450 μg上样量，等电聚焦程序重复数次除盐步骤，获得的2-DE图谱效果较好；同时对实验中遇到的常见问题进行了讨论分析。结果表明，通过优化的双向电泳条件，建立了较高分辨率和较好重复性的绒山羊皮肤组织双向电泳图谱，可以进行后续实验。本研究获得良好的绒山羊皮肤组织蛋白双向电泳图谱，验证了绒山羊蛋白质组学研究的可行性，为后续进行绒山羊皮肤毛囊发育周期蛋白质组学研究、揭示毛囊发生发育规律提供了重要的技术资料。
Abstract：To establish two-dimensional gel electrophoresis methods(2-DE) for cashmere goat skin, the experimental conditions of 2-DE were optimized, and various extraction and purification methods, different loading quantities and pH IPG strips, different isoelectric focusing were studied. The results showed that the process of grinding in liquid nitrogen and following by ultrasonication, the effect of sample purification with 2-D clean up kit, pH 4~7 17 cm pH IPG strip, the 450 μg loading quantity, isoelectric focusing process repeated several times in addition to salt step, maps were of good quality. Meanwhile, the common problems occurring in 2-DE experience were analyzed. The results indicated that the 2-DE maps of cashmere goat skin were obtained with high resolution and reproducibility, and can be used for the further proteome analysis. This study got a good 2-DE maps for cashmere goat skin protein, proved that it was feasible to conduct proteomics research for cashmere goat, and provided important technical information for researching development cycles and revealing origin and development law of skin hair follicle.
