论文部分内容阅读
本研究以向日葵幼苗为材料,比较TCA-丙酮提取法、Tris饱和酚提取法以及水溶法三种提取蛋白的方法,并且对双向电泳技术进行研究,初步建立向日葵幼苗蛋白的双向电泳体系。研究结果表明,用Tris饱和酚提取法提取蛋白含量最高,经蛋白定量后显示含量可达43.359 ug/u L;用不同浓度分离胶进行SDS-PAGE凝胶电泳,结果显示用8%分离胶可分离条带数较多,最多可分离20条蛋白条带;用不同聚焦时间以及不同上样量进行双向电泳,结果显示一向IEF聚焦时间为6 h左右,上样量为1 mg的条件较为适合,用Image Master5.0软件分析检测到蛋白点为102个。
In this study, sunflower seedlings were used as materials to compare three extraction methods of TCA-acetone extraction, Tris-saturated phenol extraction and water-soluble extraction. The two-dimensional electrophoresis technique was also studied to establish a two-dimensional electrophoresis system for sunflower seedling protein. The results showed that the content of protein extracted by Tris-saturated phenol was the highest, and the protein content was 43.359 ug / uL after protein quantification. SDS-PAGE gel with different concentrations of separation gel showed that 8% The results showed that the IEF focusing time was about 6 h, and the loading amount of 1 mg was suitable for the two-dimensional electrophoresis with different focusing time and different loading amount , Using Image Master5.0 software analysis detected 102 protein spots.