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C-乳清是胶乳的主要组分,就双向凝胶电泳技术而言,目前仍无有效的蛋白提取方法,这严重制约了双向凝胶电泳技术在橡胶树蛋白组分析上的应用。作者选取3种方法提取C-乳清蛋白,通过对蛋白产量和提取蛋白的双向凝胶电泳图谱分析系统评价3种方法。就蛋白产量而言,三氯乙酸/丙酮沉淀法产量最高(0.86mg/mlC-乳清),三氯乙酸沉淀法次之(0.72mg/mlC-乳清),酚提取法最低(0.66mg/mlC-乳清)。双向凝胶电泳图谱分析表明:酚提取法可检测到447个蛋白点,图谱背景较暗且横纵向纹理多;三氯乙酸沉淀法可检测到821个蛋白点,图谱背景居中;三氯乙酸/丙酮沉淀法可检测到1052个蛋白点,图谱背景清晰且基本无纹理。综上所述,三氯乙酸/丙酮沉淀法最适合C-乳清蛋白提取。作者首次系统评价C-乳清蛋白提取方法,必将推动橡胶树C-乳清蛋白研究发展。
C-whey is the main component of latex. For two-dimensional gel electrophoresis, there is still no effective protein extraction method, which severely restricts the application of two-dimensional gel electrophoresis in the analysis of rubber tree proteome. The authors selected three methods to extract C-lactalbumin, three methods were systematically evaluated by two-dimensional gel electrophoresis of protein yield and protein extraction. In terms of protein yield, the yield of trichloroacetic acid / acetone precipitation was the highest (0.86 mg / ml C-whey), the second was trichloroacetic acid precipitation (0.72 mg / ml C-whey) mlC-whey). Two-dimensional gel electrophoresis analysis showed that phenolics could detect 447 protein spots with darker background and more vertical and horizontal veins. 821 protein spots were detected by trichloroacetic acid precipitation method and the background of the chromatogram was centered. Trichloroacetic acid / Acetone precipitation method can detect 1052 protein spots, the background of the map is clear and almost without texture. In summary, trichloroacetic acid / acetone precipitation is the most suitable C-whey protein extraction. The authors first systematic review of C-whey protein extraction methods, will promote rubber tree C-whey protein research and development.