绵羊精子冻融前后差异蛋白质组学探究.pdfVIP

中国农业科学 2011,44(24):5092-5099 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2011.24.015 绵羊精子冻融前后差异蛋白质组学研究 李鸿岩，赵兴绪，张 勇，贺钰烜，杨永新 （甘肃农业大学动物医学院，兰州 730070 ） 摘要： 【目的】检测和分析绵羊精子冻融前后的差异蛋白质。【方法】用二维凝胶电泳分离精子蛋白，凝胶 用考马斯亮蓝染色，用 PDQuest8.0 软件检测分析差异蛋白质斑点并经液相色谱串联离子阱质谱鉴定。【结果】鲜 冻精凝胶图谱分析后得到 14 个差异蛋白斑点，在国际生物信息学中心数据库搜索到相对应的 5 种蛋白质：推定 的热休克蛋白70.1、转录因子 AP-2α蛋白、烯醇酶、烯醇酶1和血清白蛋白前体。在解冻精子中，推定的热休克 蛋白 70.1、烯醇酶和烯醇酶 1 含量减少，转录因子 AP-2α蛋白含量增加，血清白蛋白前体来源于精浆。【结论】 绵羊精子经冷冻、解冻后,蛋白质存在差异，对差异蛋白质的研究有利于探索绵羊精子在冷冻过程中的生理状态， 为进一步研究绵羊精子蛋白质组学及揭示精子冻伤机理提供依据。 关键词：绵羊；精子蛋白质；二维凝胶电泳分析；蛋白质组 Analysis of the Differential Proteomics Between Fresh and Frozen-Thawed Sheep Sperm LI Hong-yan, ZHAO Xing-xu, ZHANG Yong, HE Yu-xuan, YANG Yong-xin (College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070) Abstract: 【Objective 】To investigate the differential proteins between fresh and frozen-thawed sheep sperm. 【Method 】The current research presents the protein changes using two-dimensional gel electrophoresis (2-DE) after stained with commassie blue, differential proteins were detected by PDQuest 8.0 software and subjected to ion trap mass spectrometer equipped with a Surveyor HPLC system, and differential spots of protein were identified. 【Result 】A search against protein sequences in the National Center for Biotechnology Information databases indicated that the differential fourteen proteins correspond to five proteins including putative heat shock protein 70.1, transcription factor AP-2-alpha, enolase, enolase1 and serum albumin precursor that protein content of putative heat shock protein 70.1, enolase, enolase1 were decreased and protein content of transcript