中文摘要：

目的：建立一套适合梅花鹿鹿茸蛋白质组学研究的双向电泳方法,并以此为基础比较不同部位梅花鹿鹿茸的蛋白图谱差异。方法：使用含7 mol/L尿素、4%CHAPS、2 mol/L硫脲、65 mmol/L DTT、1 mmol/L PMSF和0.2%Bio-Lyte的溶液为蛋白裂解液提取鹿茸不同部位的总蛋白,采用固相pH 3～10线性梯度,7 cm胶条进行第一向聚焦,并对等电聚焦参数进行优化,12%SDS聚丙烯凝胶进行第二向电泳,得到的凝胶经考马斯亮蓝染色后用PDQuest软件进行图谱比较分析。结果：3个部位总蛋白含量及蛋白点数从上到下依次降低,将3个图谱经软件分析获得18个差异蛋白点,同时初步得到每个蛋白点的等电点、分子量以及灰度值。结论：优化了适合梅花鹿鹿茸蛋白的双向电泳方法;不同部位鹿茸蛋白双向电泳图谱存在明显差异。

英文摘要：

Objective ： To optimize the two-dimensional electrophoresis （2-DE） method for the proteome analysis of the Cervus nippon antler, and compare the protein maps of different parts of Cervus nippon antler. Methods：The total proteins of Cervus nippon antler were extracted by protein lysate containing 7 mol/L Urea,4% CHAPS,2 mol/L Thiourea,65 mmol/L DTT, 1 mmol/L PMSF and 0. 2%Bio-Lyte. The proteins were separated by immobilized pH 3 - 10 linear gradient（IPG） , 7 cm strips as the first dimension. Isoelectric fo- cusing conditions were optimized. 12% SDS-PAGE was used as the second dimension electrophoresis. The gels were stained with Coomassie brilliant blue and analyzed by PDQuest analysis software. Results：The contents of total protein and the numbers of protein points of three different parts of Cervus nippon antler reduced gradually from the top to the bottom. Comparing three maps of different parts of Cer- vus nippon antler, there were 18 different protein points. Isoelectric point, molecular weight and gray value of each different protein point were calculated. Conclusion：An optimized two-dimensional electrophoresis method for the proteome analysis of the Cervus nippon antler is established. The 2-DE profiles of different parts of Cervus nippon antler exist obvious differences. The different protein points can be used as reference for Cervus nippon antler quality control and evaluation.