西花蓟马蛋白的提取及双向电泳体系的建立

【目的】蛋白样品的制备是获得良好双向凝胶电泳(2-DE)图谱的前提,建立合理的西花蓟马蛋白的双向电泳体系,获得分辨率较高、重复性较好的图谱,能够为后续的研究提供有力支撑。【方法】实验以西花蓟马成虫为实验材料,对比了饱和酚法、TCA/丙酮法和直接裂解法3种蛋白提取方法,从中选出最适宜双向电泳分析的一种蛋白提取方法。【结果】3种方法蛋白提取率差异显著,直接裂解法蛋白提取率最高,饱和酚法的蛋白提取率最低;3种方法的SDS-PAGE条带数差异不明显;TCA/丙酮法的双向凝胶图谱效果最好,蛋白点最多。【结论】TCA/丙酮法能够有效去除西花蓟马蛋白中的干扰物质,是最适合西花蓟马双向凝胶电泳的蛋白提取方法,为后续西花蓟马在蛋白组学方面的研究奠定了基础。

Protein extraction of Frankliniella occidentalis and establishment of two-dimensional electrophoresis system

[Aim] Protein sample preparation is the prerequisite to obtain an optimal two-dimensional electrophoresis (2-DE) gel map. Establishing a suitable 2-DE system can provide a strong support for the subsequent research.[Method] This study used Frankliniella occidentalis adults as experimental materials, and compared three protein extraction methods:Tris-phenol, TCA/acetone precipitation, and direct decomposition extraction. The most suitable method of 2-DE system was defined through the analysis of 2-DE protein images.[Result] Experimental results revealed that the three methods showed significant differences in protein extraction rate, among which the direct decomposition method was the highest while Tris-phenol method was the lowest. The three methods got similar SDS-PAGE results. TCA/acetone method had the best quality of 2-DE gel map, with the highest number of protein spots.[Conclusion] The results suggest that TCA/acetone method could remove interfering substance effectively and was optimal for protein extraction of F. Occidentalis. It laid foundation of further study on the proteomics of F. occidentalis.