健康奶牛与临床型乳腺炎奶牛乳腺核蛋白组差异表达分析  

Proteomic Analysis of Nuclei of Mammary Tissue from Healthy Cows and Clinical Mastitic Cows

作　　者：王建锋[1] 赵兴绪[1] 张勇[1] 徐铁山[1] 

机构地区：[1]甘肃农业大学动物医学院,兰州730070

出　　处：《畜牧兽医学报》2010年第1期105-111,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：科技部国际合作重点项目计划(2005DFA30720)

摘　　要：亚细胞蛋白质组的优点在于对低丰度蛋白的研究,运用亚细胞蛋白质组学的研究策略,可以提高低丰度蛋白质在双向电泳中的检出数量。通过分析奶牛乳腺炎乳腺与正常乳腺核蛋白组差异表达情况,为奶牛乳腺炎发病机理研究寻找尽可能多的生物标记分子。经超速离心法分离细胞核,双向凝胶电泳分离蛋白,用PDQuest7.4软件分析寻找差异蛋白质斑点,高效液相色谱串联离子阱质谱鉴定蛋白质。从核蛋白组2-DE图谱中筛选出22个差异表达的蛋白质斑点,质谱鉴定出17个差异表达的蛋白质,2个蛋白质在乳腺炎发病过程中下调,7个上调,5个只在正常情况下表达,3个只表达在乳腺炎组织中,筛选出的差异表达蛋白质涉及细胞骨架构成、代谢调节及凋亡调控等许多方面。表明奶牛乳腺炎发生时乳腺组织核结构和代谢状态都发生了的变化。The sub-cellular proteomics play an important role in study of minor proteins,which participating the intracellular metabolism.Sub-cellular proteomics can increase the minor proteins separation by two-dimensional electrophoresis（2-DE）.A comparative analysis of proteomic profiling was conducted between nuclei of healthy mammary gland and infected clinical mastitis of dairy cow,to find more valuable biological molecular labels involved in mastitis pathogenesis.In this study,nuclei were isolated and purified by ultracentrifuge,and then two-dimensional electrophoresis was applied to separate nuclear proteins.Imagine analysis for selection of differentially-expressed protein spots was conducted by PDQUest 7.4 software.By ion trap mass spectrometry equipped with surveyor HPLC system proteins were identified.22 spots from electrophoresis gels were selected as differentially-expressed proteins for MS analysis and 17 proteins were identified.Seven of differentially-expressed proteins showed upregulated expression and 2 proteins showed down-regulated expression during mastitis.5 proteins only expressed in healthy mammary gland and 3 proteins only expressed in mammary gland which infected with clinical mastitis.The differentially-expressed proteins separated in this study had a wide range of functions which are related to the formation of cellular skeleton,metabolic regulation and apoptosis regulation.These suggested that nuclei of mammary gland had experienced fundamental changes in structure and metabolism when contracted clinical mastitis.

关 键 词：奶牛 乳腺 细胞核 亚细胞蛋白质组 

分 类 号：Q51[生物学—生物化学]