甘蓝型油菜蛋白质双向电泳体系的建立  

Construction of Protein Two-Dimensional Polyacrylamide Gel Electrophoresis System for Brassica napus

作　　者：甘露[1,2] 李殿荣[1,3] 臧新[4] 付春华[1,2] 余龙江[1,2] 栗茂腾[1,2] 

机构地区：[1]华中科技大学生命科学与技术学院,湖北武汉430074 [2]分子生物物理教育部重点实验室,湖北武汉430074 [3]陕西省杂交油菜研究中心,陕西大荔715105 [4]郑州大学生物工程系,河南郑州450001

出　　处：《作物学报》2010年第4期612-619,共8页Acta Agronomica Sinica

基　　金：国家高技术研究发展计划(863计划)项目(2009AA101105);新世纪优秀人才支持计划;华中科技大学自主创新基金(M2009059)项目资助

摘　　要：选用油菜品种08127,采用TCA-丙酮提取方法,pH4～7胶条和改进的IEF聚焦程序,得到较好的蛋白质双向电泳结果,建立了一种适合甘蓝型油菜的双向电泳体系。利用其他甘蓝型油菜品种的幼苗、叶片和茎中的蛋白检验该实验体系,都能得到较好的蛋白质双向电泳结果。利用该实验体系比较甘蓝型油菜幼苗不同发育时期蛋白质双向电泳图,找到二者之间的差异点,选取部分点成功进行了质谱分析。Two-dimensional polyacrylamide gel electrophoresis has been widely applied in proteomic researches. The approach can analyze dynamic changes of proteomics of biological samples under different conditions to identify specific proteins and genes. The two-dimensional gel systems for rice,Arabidopsis,and other species have been established. The purpose of this re-search was to establish a suitable system for B. napus to analyze proteomics. Rapeseed variety of 08127 was used in the experi-ment,the proteins were obtained by the trichloroacetic extraction method with pH 4–7 gel brands under the improved IEF procedure,showing a clean two-dimensional electrophoresis map. Therefore,a proper system for B. napus was formed. Then the system was validated by using the young seedlings,leaves and stems from other varieties,and the same results as those from variety 08127 were obtained. The proteomics profiling of seedlings in different ages of B. napus was analyzed by using the previous established protocol,we found 20 up-regulated and 51 down-regulated proteins with more than two times changes in 30 days old B. napus stems compared with in 15 days old B. napus stems and some of the different protein spots were successfully identified. The system established in this study is useful for B. napus proteomic research in the future.

关 键 词：甘蓝型油菜 蛋白质 双向电泳 

分 类 号：S565.4[农业科学—作物学]