SDS-PAGE与液质联用技术分离和鉴定中国明对虾(Fenneropenaeus chinensis)血浆蛋白

PROFILING THE PLASMA PROTEOME OF FENNEROPENAEUS CHINENSIS WITH SDS-PAGE AND LC-MS/MS

提取中国明对虾的血浆蛋白用SDS-PAGE进行分离。将电泳条带从上到下平均切成4份,分别胶内酶解并利用液相色谱-串联质谱系统进行质谱分析。利用SEQUEST软件对NCBInr蛋白数据库进行检索,鉴定了66个中国明对虾血浆蛋白质。运用生物信息学手段,对鉴定的蛋白进行功能分类,包括转录、细胞结构和骨架、信号转导、蛋白质合成与处理、蛋白质折叠、分拣和降解、免疫功能、生长和发育、能量代谢、细胞周期、碳水化合物代谢、钙离子平衡、抗氧化剂和氨基酸和脂类代谢等。本研究丰富了中国对虾血浆蛋白质表达谱,可为进一步研究对虾免疫机制提供有力的支持。

Plasma, as a major component of hemolymph, plays important physiological functions in Chinese shrimp Fenneropenaeus chinensis immunity. However, it is difficult to analyze plasma proteome by 2-DE because plasma is a complicated and dynamically changing system, consisting of different types and abundant proteins. In this study, SDS-PAGE pre-fractionation combined with LC-MS/MS （liquid chromatography-tandem mass spectrometry） was applied to achieve more comprehensive proteome profiles of plasma proteins. 66 non-redundant proteins were identified in this research. By means of bioinformatics, the identified proteins was classified in terms of functions in transcription, structural and cytoskeletal, signal transduction, protein synthesis, protein folding-sorting-degradation, immunity, growth performance, energy metabolism, cell cycle, carbohydrate metabolism, calcium homeostasis, antioxidant and amino acid, and lipid me- tabolism, which shall enhance our understanding on the immunity mechanisms of Chinese shrimp.