| 叶用莴苣叶片总蛋白质双向电泳影响因素的研究 |
| |
| 引用本文： | 陈曙红,韩莹琰,范双喜,李洋. 叶用莴苣叶片总蛋白质双向电泳影响因素的研究[J]. 北京农学院学报, 2012, 27(2): 14-16 |
| |
| 作者姓名： | 陈曙红 韩莹琰 范双喜 李洋 |
| |
| 作者单位： | 北京农学院植物科学技术学院,北京,102206;北京农学院植物科学技术学院,北京,102206;北京农学院植物科学技术学院,北京,102206;北京农学院植物科学技术学院,北京,102206 |
| |
| 基金项目： | 北京市自然科学基金，北京市教育委员会科技计划重点项目，北京市自然科学基金，北京市科技新星计划项目，北京市属高等学校人才强教计划资助项目 |
| |
| 摘 要： | 以叶用莴苣叶片为试材,研究17cm固相pH梯度胶条(pH 4～7)体系下双向电泳不同的提取方法、上样体积、定量方法等影响因素,力求确定其理想的双向电泳条件。结果表明,酚-甲醇/醋酸铵沉淀法对蛋白质提取更彻底,效果更好。对于蛋白质定量的试验,相比于常规的Bradford方法,利用无干扰蛋白质定量试剂盒具有更好的准确性。考虑叶片总蛋白质中大量Rubisco的存在,应相对增加上样量,采用考马斯亮蓝染色时取800μg到1 100μg蛋白质进行双向电泳比较合理。
|
| 关 键 词： | 叶用莴苣 叶片 蛋白质提取方法 双向电泳技术 |
Study on the Influential Factors of 2-Dimensional Electrophoresis in Lettuce Leaf |
| |
| CHEN Shu-hong , HAN Ying-yan , FAN Shuang-xi , LI Yang. Study on the Influential Factors of 2-Dimensional Electrophoresis in Lettuce Leaf[J]. Journal of Beijing Agricultural College, 2012, 27(2): 14-16 |
| |
| Authors: | CHEN Shu-hong HAN Ying-yan FAN Shuang-xi LI Yang |
| |
| Affiliation: | (College of Plant Science and Technology,Beijing University of Agriculture,Beijing 102206,China) |
| |
| Abstract: | The experiment,taking lettuce leaf as experiment material,investigated different protein extraction methods,different sample weights,different quantitative methods,and other influential factors in order to get the optimal two-dimensional electrophoresis(2-DE) gel patterns in 17cm immobilized pH gradients strip(pH4~7) system.The phenol extraction-methanol/ammonium precipitation extraction method showed a good effect.Compared to the normal Bradford quantitative method,the non-interfering protein quantitative assay kit was more accurate for protein quantitative experiment.Considering too much Rubisco in the total protens,loading weight of sample protein should be increased,and the optimal loading weight was about 800μg-1100μg. |
| |
| Keywords: | Lettuce(Lactuca Sativa L.) Leaf Protein extraction method 2-DE |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
