CAJ | 학술논문

高质量提取大白菜花蕾蛋白是大白菜花器官蛋白质组学研究的关键步骤.本试验以大白菜AB01可育花蕾为材料,采用TCA丙酮沉淀法、Tris-HC1法、酚改良法、Trizol沉淀法、Tris-丙酮-酚法和尿素-硫脲提取法等六种方法分别提取花蕾全蛋白,通过Bradford法和双向电泳蛋白定量试剂盒测定蛋白浓度,经双向电泳分离后得到2-DE图谱,经分析、比较图谱蛋白点的情况,找到花蕾全蛋白的最佳提取方法和浓度测定方法.结果显示:TCA丙酮沉淀法得到的2-DE图谱,蛋白点分布均匀、清晰,是一种较为理想的花蕾蛋白提取方法;采用蛋白定量试剂盒测定蛋白浓度更加准确.
고질량제취대백채화뢰단백시대백채화기관단백질조학연구적관건보취.본시험이대백채AB01가육화뢰위재료,채용TCA병동침정법、Tris-HC1법、분개량법、Trizol침정법、Tris-병동-분법화뇨소-류뇨제취법등륙충방법분별제취화뢰전단백,통과Bradford법화쌍향전영단백정량시제합측정단백농도,경쌍향전영분리후득도2-DE도보,경분석、비교도보단백점적정황,조도화뢰전단백적최가제취방법화농도측정방법.결과현시:TCA병동침정법득도적2-DE도보,단백점분포균균、청석,시일충교위이상적화뢰단백제취방법;채용단백정량시제합측정단백농도경가준학.
@@@@Extracting high-quality flower bud proteins is the prerequisite in studying of the floral-organ Proteome in Chinese cabbage. In this experiment, the flower buds of Chinese cabbage A/B line AB01 were used to extract the whole protein by using the following six methods (TCA acetone precipitation method, Tris-HC1 method, Phenol modified method, Trizol precipitation method, Tris-acetone-phenol method and Urea-thiourea extraction method). The 2-DE maps were generated by the two-dimensional electrophoresis approach as well. The best methods for flower bud protein extraction and concentration determination were selected by analyzing and comparing the protein spots in 2-DE maps. The results showed that the protein spots in the 2-DE maps generated by TCA-acetone precipitation were clear and even distributed, therefore the TCA-acetone precipitation method was recommended as an ideal methods for extracting the flower bud protein, while the Protein Assay kit should be more accurate to detect the protein concentration based on comparing the maps generated by the two-dimensional electrophoresis protein quantitative kit and Bradford method.