Abstract: [Objective] To establish two-dimensional gel electrophoresis methods (2-DE) for sheep serum.[Method]The experimental conditions of 2-DE were optimized,and various extraction and process methods,different pH IPG strips,different concentration of SDS-PAGE were studied.[Result] The results showed that the 10; tca/ acetone precipitation method was used to deal with the serum protein,pH 4-7 IPG,2.5 mg,18 cm,10; sds-page,and a good 2-DE map was obtained.[Conclusion]This study got a good IPG maps for sheep serum protein.We have successfully established a 2-DE technique for sheep serum proteome,which can provide important technical information for evaluation of transgenic sheep biological safety.

摘要： [目的]建立和优化绵羊血清蛋白双向电泳体系(2-DE).[方法]从样品提取及处理、IPG胶条的选择、SDS-PAGE浓度的选择等多个方面对双向电泳分离条件进行比较.[结果]采用10; TCA/丙酮沉淀法处理血清蛋白质,18 cm pH 4～7的IPG胶条2.5 mg上样量,10; SDS-PAGE,获得的2-DE图谱效果较好.[结论]获得良好的绵羊血清蛋白双向电泳图谱,建立并优化了绵羊蛋白质组学的双向电泳技术体系,为转基因绵羊生物安全性评价提供了重要的技术条件.