雏鸡冷应激前后血清中差异蛋白质组学研究

本试验旨在比较分析雏鸡冷应激前后血清中蛋白质的表达差异,并对重点差异蛋白质进行鉴定。将30只雏鸡随机分为3组,分别为冷应激组、冷适应组和常温对照组,收集各组雏鸡的血液制备血清后进行双向凝胶电泳(2-DE),以获得血清蛋白质表达的2-DE图谱,对2-DE图谱进行差异分析(利用PDQuest 8.0软件),随后采用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术对差异表达蛋白质进行鉴定,并利用蛋白质印迹(Western blot)方法进行验证。结果显示:通过2-DE对常温对照组、冷应激组和冷适应组雏鸡血清进行分析,得到了比较完整的差异蛋白质数据,共找到差异蛋白质点23个。采用MALDI-TOF-MS技术分析其中几个重复性好且较为明显的蛋白质点,成功鉴定出4个差异蛋白质,其中2个为果糖二磷酸醛缩酶C(ALDOC),是参与葡萄糖、能量代谢通路供能的相关蛋白;随后,对差异蛋白质ALDOC进行Western blot验证,所得结果与2-DE的结果相一致。结果表明,雏鸡冷应激前后血清中蛋白质的表达具有明显的差异,这些蛋白质的表达差异可能与冷应激有关。

A Study on Differential Proteomics of Chicks Serum before and after Cold Stress

The objective of this experiment was to compare and analysis the difference of protein expression in serum of chicks before and after cold stress, and to identify the differential proteins. Thirty chicks were ran?domly divided into three groups, which were cold stress group, cold?adapted group and normal temperature control group. Blood samples of chicks in each group were collected and prepared the serum for two?dimension?al gel electrophoresis ( 2?DE) , in order to obtain the 2?DE maps for protein expressions in serum. The differ?ences of 2?DE maps were analysed by PDQuest 8.0 software. After chicks' serum differential expression pro?teins were analysed, matrix?assisted laser desorption ionization time of flight mass spectrometry ( MALDI?TOF?MS) technology was used to identify the differential expression proteins, and the differential expression pro?teins were verified by Western blot method. The results revealed that a relatively complete differential protein data for chicks' serum in normal temperature control group, cold stress group and cold?adapted group were ob?tained by 2?DE analysis, and 23 differential protein spots were detected. MALDI?TOF?MS technology was uti?lized to analyze several of reproducible and more obvious protein spots, successfully identifying 4 different pro?teins, two of them were aldolase C fructose?bisphosphate ( ALDOC) , which was related protein involved in glucose and energy metabolism pathway. Then, by using the Western blot method for further validated differen?tial protein ALDOC, the results were consistent with the results of the 2?DE. Therefore, the results indicated that the expressions of proteins in serum of chicks before and after cold stress are significantly different. The ex?pression differences of these proteins may relate to cold stress.