绵羊血清蛋白双向凝胶电泳技术的建立

Establishment of Two- dimensional Gel Electrophoresis Technique for Sheep Serum

【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 p H 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。

[ Objective] To establish two- dimensional gel eleetrophoresis 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 rag, 18 era, 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.