动物营养学报 >

2017 , Vol. 29 >Issue 5: 1567 - 1573

DOI: https://doi.org/10.3969/j.issn.1006-267x.2017.05.015

分子营养 Molecular Nutrition

雏鸡冷应激前后血清中差异蛋白质组学研究

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  • 黑龙江八一农垦大学动物科技学院, 大庆 163319
杨玉英(1967-),女,黑龙江海伦人,教授,硕士,从事预防兽医学研究。E-mail:yalele258@sina.com

收稿日期: 2016-11-11

  网络出版日期: 2017-05-13

基金资助

十三五国家重点研发计划（2016YFD0501210）；国家自然科学基金面上项目（31272524）；农业部948计划重点项目（2011-G35）

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

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  • College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China

Received date: 2016-11-11

  Online published: 2017-05-13

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摘要

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。

关键词： 双向电泳; 冷应激; 蛋白质组; 蛋白质表达差异; 差异蛋白质点

本文引用格式

杨玉英, 刘鹏, 贺军, 刘洋, 姚睿智, 史宏昭, 甄莉, 唐德江, 李士泽 . 雏鸡冷应激前后血清中差异蛋白质组学研究[J]. 动物营养学报, 2017 , 29(5) : 1567 -1573 . DOI: 10.3969/j.issn.1006-267x.2017.05.015

Abstract

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 randomly 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-dimensional gel electrophoresis (2-DE), in order to obtain the 2-DE maps for protein expressions in serum. The differences of 2-DE maps were analysed by PDQuest 8.0 software. After chicks' serum differential expression proteins 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 proteins 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 obtained by 2-DE analysis, and 23 differential protein spots were detected. MALDI-TOF-MS technology was utilized to analyze several of reproducible and more obvious protein spots, successfully identifying 4 different proteins, 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 differential 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 expression differences of these proteins may relate to cold stress.

Key words： two-dimensional electrophoresis; cold stress; proteome; differential expression protein; differential protein spots

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