猪卵泡液样品制备方法对双向电泳图谱的影响  

Methods of FF Sample Preparation on the Effect of Map Quality of Two-Dimentional Electrophoresis in Sows

作　　者：孙艳玲[1] 周虚[1] 李晓艳 王兴龙 李成娇[1] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]军科院军事兽医研究所,长春130062

出　　处：《生物技术通报》2010年第5期149-153,157,共6页Biotechnology Bulletin

基　　金：国家科技支撑项目(2006BAD14B08-01);国家自然科学基金项目(30671511)

摘　　要：比较不同的蛋白制备方法,寻找适合猪卵泡液的双向电泳样品制备方法,提高2-DE图谱的分辨率和重复性。利用ProteoExtract Albumin/IgG Remove Kit去除卵泡液中的高峰度蛋白;然后分别用丙酮沉淀法、TCA/丙酮沉淀法和超滤法进行样品浓缩,比较分析对2-DE图谱的影响。结果表明,TCA/丙酮沉淀法制备的蛋白样品获得的电泳图谱质量较好,蛋白斑点数目较多,斑点较清晰、圆滑,分辨率较高,重复性较好。TCA-丙酮沉淀法更适合于制备猪卵泡液的双向电泳蛋白样品,为进一步对猪卵泡液2-DE图谱进行分析、鉴定及寻找生物标记物提供了保证。Accumulating of proline in plant subjected to adverse environment is one of the most important adaptations,which can re- duce damage on plant. Proline metabolism,and proline functions in plants encountered with adverse environments were summarized in this paper. The consentaneous conclusion indicated that the expression of P5CS gene was up-induced,and the increasing expression of proline synthetase gene could help for increasing the proline accumulation in plants,which could improve plant tolerance to adverse circumstance. The study was to determine effective methods of sample preparation of two-dimensional electrophoresis of follicular fluid in sows,to improve its resolution and reproducibility. The sample were depleted the high abundant proteins by ProteoExtract Albumin/IgG Remove Kit,and then concentrated by Acetone precipitation,TCA-acetone precipitation method and ultrafiltration method to compare the effect on electrophoresis. Sample prepared by TCA-aeetone precipitation,obtained more and clearer protein spots,and high resolution and bet- ter reproducibility of 2-DE maps than that of others. It proved that TCA-acetone precipitation is suitable for preparing the samples of follicular fluid in sows,which could provide a foundamental guarantee for further analysis of 2-DE maps and finding biomarkers.

关 键 词：卵泡液 蛋白 制备方法 双向电泳 

分 类 号：Q503[生物学—生物化学]