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目的:建立适用于酸浆种子的SDS-PAGE和2-D PAGE蛋白样品提取方法。方法:比较直接提取法、TCA-丙酮法、Tris-丙酮法、饱和酚法、改良饱和酚法等7种提取方法所获酸浆种子总蛋白含量以及SDS-PAGE电泳的谱带数目、强度和电泳分辨率等方面的差异。结果:HEPES buffer直接提取法提取蛋白效率较高,杂质少,电泳谱带清晰,分辨率高;PVP buffer和Lysis buffer A直接提取法次之,但杂质较多;TCA-丙酮法、Tris-丙酮法、饱和酚法和改良的饱和酚法差别不大,蛋白纯度较高,但提取效率低,谱带数目少;此外,Lysis buffer A法、丙酮法和酚法易产生高丰度蛋白的干扰。结论:直接提取法中的HEPES buffer法最好,优于丙酮法、酚法和其它直接提取法。
OBJECTIVE: To establish a method for the extraction of SDS-PAGE and 2-D PAGE protein samples suitable for physalis seed. Methods: The total protein content of the pulp and the number and intensity of the SDS-PAGE electrophoresis were compared by direct extraction, TCA-acetone, Tris-acetone, saturated phenol and modified saturated phenol Electrophoretic resolution and other aspects of the difference. Results: The direct extraction method of HEPES buffer was more efficient and less impurities, the bands of electrophoresis were clear and the resolution was higher. The direct extraction method of PVP buffer and Lysis buffer A was the second, but the impurities were more; the TCA-acetone method and Tris-acetone Method, saturated phenol method and modified saturated phenol method are not significantly different, the protein purity is high, but the extraction efficiency is low, the number of bands is small; In addition, Lysis buffer A method, acetone method and phenol method easy to produce high abundance protein interference . Conclusion: The HEPES buffer method in the direct extraction method is the best, better than the acetone method, phenol method and other direct extraction method.