检测肥育猪CAST蛋白的免疫印迹方法研究  

Study on Western Blot Method to Detect CAST in Growing-Finishing Pigs

作　　者：张勇[1] 傅刚[2] 翟峰[1] 朱宇旌[1] 孙景霞 邵彩梅[4] 

机构地区：[1]沈阳农业大学畜牧兽医学院,沈阳110161 [2]沈阳大学,沈阳110044 [3]大连老虎滩海洋公园鸟语林,辽宁大连116013 [4]辽宁禾丰牧业股份有限公司,沈阳110164

出　　处：《沈阳农业大学学报》2011年第3期321-324,共4页Journal of Shenyang Agricultural University

基　　金：国家自然科学基金项目(30500363)

摘　　要：为了研究3个不同蛋白质水平(10%、18%、26%)饲粮对肥育猪的钙蛋白抑制蛋白酶(CAST)的影响,对CAST蛋白提取、电泳、转膜等条件进行优化。试验表明:CAST被激活的条件是将蛋白提取液于上样缓冲液95℃煮15min后,立即在冰块上冷却,防止蛋白降解;采用分离胶浓度为10%进行电泳,可以明显检测到68kD、107 kD的CAST;采用预染Marker和DAB染色液可针对性地检测到目的蛋白的位置;利用上述程序检测到了猪CAST两种类型的表达,取得了较满意的结果。Western blotting method to investigate effects of different dietary levels protein （10%,18% and 26%） on the expressionof CAST of growing-finishing pigs was studied. Its procedures including the extraction of protein, the condition of electrophoresis and immuno-blotting were optimized. The results showed that CAST was activated after the extract was heated at 95℃ for 15min and then cooled on the ice immediately in order to prevent being degraded. CAST of 68 kD and 107 kD was detected by using a separation gel of 10% for SDS-PAGE preparation, the target protein position was detected directly by pre-stained Marker and DAB staining solution. Two types of CAST were detected successfully by the above procedures.

关 键 词：CAST 免疫印迹 肥育猪 

分 类 号：S828.91[农业科学—畜牧学]