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蛋白质组学研究的关键问题之一是双向电泳(2-DE)中低丰度蛋白的分析,主要是因为相关高丰度蛋白的存在,导致IPG胶条无法吸胀低丰度蛋白,使得低丰度蛋白在2-DE中很难被检测出来。利用梯度浓度的PEG4000沉淀方法来富集绿竹叶片中不同丰度的蛋白质,从而使得低丰度蛋白在凝胶中显现出来。在PEG分级法和传统的全蛋白提取法的2-DE比较中,发现采用PEG分级沉淀法提取的蛋白质的数量以及类别明显增加,高丰度蛋白主要富集在8%和16%的PEG浓度组分中,使得其他组分中的低丰度蛋白与高丰度蛋白组分分别进行2-DE分析。经过对图像和数据的比较、分析,PEG分级法得到的5个组分蛋白点总数超过了1032个,大约是传统蛋白提取方法制备蛋白样品的3倍。
One of the key issues in proteomics research is the analysis of low-abundance proteins in two-dimensional electrophoresis (2-DE), primarily because of the presence of related high-abundance proteins, resulting in the inability of IPG strips to upset low-abundance proteins, making low Abundance of proteins is hard to detect in 2-DE. Gradient concentration of PEG4000 precipitation method to enrich the abundance of green bamboo leaves of different abundance of protein, making low-abundance protein in the gel appear. In the 2-DE comparison of the PEG fractionation method and the traditional whole protein extraction method, it was found that the number and the categories of the proteins extracted by the PEG fractionation precipitation method were significantly increased, and the abundance of the high abundance protein mainly concentrated at PEG concentrations of 8% and 16% Components, so that low abundance of other components and high abundance protein fractions were 2-DE analysis. After comparing the images and data, the total number of protein spots of the five components obtained by the PEG grading method exceeded 1032, about three times that of the traditional protein extraction method.