大鼠附睾蛋白质组的提取和二维液相色谱分离

背景与目的：提取犬鼠附睾液蛋白并建立一种利用二维液相色谱法分离附睾蛋白组的方法。方法：分离提取犬鼠附睾液蛋白。样品利用起始缓冲液置换后，进行一维色谱聚焦分离，然后收集pH8．5—4．0之间的组份进行二维反相离压液相色谱分离，最后将获得的二维UV图通过ProteoVue软件转换成PI／UV图谱。结果：成功提取了附睾液蛋白，并通过二维液相色谱成功建立了大鼠头体尾部附睾液蛋白的二维PI／UV图谱，收集了一维色谱聚焦分离的pH8．5—4．0区间的20个组份，并将每个组份进行二维色谱分离后转换为PI/UV图谱。结论：为进一步全面研究附睾蛋白功能和体液差异蛋白质组研究打下了基础。

Extraction and two- dimensional liquid chromatographic fractionation of rat epididymal fluid proteins

Background and Objective: To extract rat epididymal fluid proteins and develop an approach for fractionating the proteins by two dimensional liquid chromatography.Methods: The epididymal fluid proteins extracted from three rats were exchanged with start buffer and separated by chromatofocusing in the first dimension,the fractions between pH8.5 and pH4.0 collected from 1D were separated by non-porous silica reverse-phase HPLC.The UV maps were transformed into PI/UV maps by ProteoVue software.Results: We successfully extracted epididymal fluid proteins and 20 fractions between pH8.5-pH4.0 were collected,each fraction was separated by 2D and the UV maps were transformed into PI/UV maps.Conclusion: Our study laid a foundation for further studies of epididymal fluid proteins and other body fluid Differential Proteome.