【摘要】 【目的】通过分析临床健康和乳房炎奶牛乳腺组织膜蛋白的差异表达,以期在蛋白质水平探索奶牛乳房炎的发生机理。【方法】采用二维凝胶电泳技术分离了临床健康和乳房炎奶牛的乳腺组织膜蛋白,经考马斯亮蓝染色后PDQuest 7.4软件匹配、检测凝胶中差异表达的蛋白点,对12个差异蛋白点采用高效液相色谱串联离子阱质谱分析,SEQUEST软件搜索NCBInr数据库。半定量RT-PCR分析差异表达蛋白的mRNA水平。【结果】12个差异蛋白点鉴定为11种蛋白质,其中9个蛋白点在临床型乳房炎奶牛乳腺组织膜蛋白中表达量上调而3个蛋白点表达量下调,主要涉及细胞膜骨架系统、信号转导、物质结合和传递等生物学功能。半定量RT-PCR分析表明,乳房炎奶牛乳腺组织中钙粒蛋白B的mRNA表达水平是健康奶牛的1.6倍。【结论】推测这些差异蛋白的表达变化与奶牛乳房炎发生相关,为从蛋白质水平揭示奶牛乳房炎的发生机理以及发现潜在的治疗目标蛋白提供了理论依据。更多还原

【Abstract】 Objective The objective of the paper was to investigate different mammary membrane proteins between clinical healthy dairy cows and dairy cows with mastitis,and to further for exploring the mechanism of pathogenesis.Method This study presented the membrane protein changes of mammary gland from clinical healthy dairy cows and dairy cows with mastitis,using two-dimensional gel electrophoresis(2-DE).After the protein stained with Coomassie Blue G-250,twelve differential expression protein spots were detected by PDQuest 7.4 software and then subjected to the HPLC coupled with ion trap mass spectrometer system.Proteins were identified using the SEQUEST software to search the NCBInr database,and mRNA level of differential expression protein was presented by semi-quantity RT-PCR.Result Eleven proteins were identified.Three proteins were down-regulated and eight proteins were up-regulated in cows infected with clinical mastitis,they were involved in signal transduction,binding and transport activity.The mRNA level of calgranulin B expression in mammary gland of cows with clinical masitis was 1.6-fold higher than the healthy dairy cows confirmed by semi-quantity RT-PCR protocol.Conclusion The results may provide valuable information for the investigation of the host mammary immune system responses to defense against pathogens at the protein level and potential protein targets for treatment.更多还原