蚯蚓蛋白质组学研究中3种总蛋白提取方法的比较  

Comparison of Three Total Protein Extraction Methods in Earthworm Proteomics Research

作　　者：郭锦[1,2] 高宏生[1] 赵宏[4] 杨震[3] 张丽[1] 胡骁[1] 李华英[1] 赵化冰[1] 

机构地区：[1]中国人民武装警察部队后勤学院,天津市职业与环境危害防制重点实验室,天津300309 [2]南开大学环境科学与工程学院教育部环境污染过程与基准重点实验室,天津300071 [3]中国人民武装警察部队后勤学院,附属医院科研科,天津300162 [4]天津出入境检验检疫局动食植检测中心,天津300461

出　　处：《南开大学学报（自然科学版）》2016年第1期1-6,共6页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：国家自然科学基金重点项目(21037002);国家自然科学基金(21477163);天津市应用基础与前沿技术研究计划重点项目(14JCZDJC40400)

摘　　要：双向电泳(2-DE)技术在现阶段的蛋白质组学研究当中,由于其价格相对便宜,仪器设备相对简单而被广泛应用.合适的总蛋白质样品制备方法,可以提高双向电泳蛋白点的数目和分辨率以保证蛋白质组信息的完整性,是整个实验的关键.蚯蚓作为土壤污染和土壤生态毒理学研究的重要生物,其总蛋白质提取的方法学研究还不完善.通过对已发表的文献中的蛋白提取方法进行筛选和优化,利用SDS-PAGE电泳和双向电泳的结果,对三氯乙酸-丙酮沉淀法、裂解液法和酚抽提法对蚯蚓总蛋白提取结果进行了评价,建立了适合蚯蚓蛋白质组学研究的总蛋白样品制备方法.Two- dimensional gel electrophoresis technique has been widely used for proteomic studys due to its relatively simple operation and acceptable price. The proteins would be separated and fractioned by their p I and MW for the subsequent identification by mass spectrum. As a result, a suitable sample preparation method may increase the number of protein spots and the resolution of different proteins, which are significant for the integrity of proteome information and it is the key step in the whole research. How to extract the total proteins as many as possible plays an important role in sample preparation. Because of the difference among various oranisms, tissues and cells, there is no versatile and effective extraction method for diverse samples. A sample-specific extracttion protocol should be searched by some comparative experiments. In this study, we screened and optimized from several reported protein extraction protocols and the extraction effects of Eiseina fetida total proteins were evaluated by SDS-PAGE and 2-DE. Finally we established a better extraction protocol and laid the foundation for the future proteomic study on earthworm.

关 键 词：双向电泳 三氯乙酸 赤子爱生蚯蚓 样品制备 

分 类 号：TQ93[化学工程]