作　　者： ; ; ; ;

机构地区： 湖南师范大学生命科学学院

出　　处： 《广东海洋大学学报》 2007年第3期117-122,共6页

摘　　要： 为找寻光温敏核不育水稻幼穗与育性调控有关的蛋白质,采用固相pH梯度-SDS聚丙烯酰胺双向凝胶电泳对光温敏核不育水稻(培矮64S)育性转换期幼穗总蛋白进行了分离,获取凝胶图像并进行重复性分析,通过微量测序-Edman降解方法和原位酶解及MALDI-TOF质谱的方法鉴定了60个蛋白质点。结果表明:本方法可得取分辨率和重复性较好的凝胶图谱。蛋白质点在2D胶上的重复性为:沿等电聚焦方向偏差为1.49±0.22 mm,沿SDS-PAGE方向偏差为1.24±0.18 mm。37个蛋白质点得到了准确的鉴定结果,其中16个是差异蛋白质点。在不育变化为可育的过程中,明显表达上调的蛋白质点包括几丁质酶,丙酮酸脱氢酶等;明显下调的蛋白质包括超氧化物歧化酶,硝酸还原酶脱辅基酶蛋白等;金属硫蛋白RicMT只在可育中存在。 The proteins of photo-thermo-sensitive genetic mail sterile rice（ORYZA SATIVA, peiai64S） spikelet on fertility alternation stage were separated by two-dimensional electrophoresis with immobilized pH （3-10） gradients as the first dimension and SDS-PAGE as the second. 2-D maps with high quality and reproducibility were obtained. Mean standard deviations of 1.49± 0.22 mm for IPG-IEF and 1.24±0.18 mm for SDS-PAGE were obtained. 60 protein spots out of about 1000 detectable spots on the 2-D gels were indentified by the following two methods： 1）N- terminal Edman degradation microsequencing after the protein spots were electro-transferred to PVDF membrane. 2）MALDI-TOF-MS peptide fingerprint analysis of the protein spots and protein database searching. The 2D protein maps of the rice spikelet during the sterile and fertile stages were compared. While the rice spikelet changed from sterile to fertile stages,about 16 protein spots showed variations that were significant and reproducible. Six up-regulated proteins and nine down-regulated proteins were identified. One protein only exist in fertile stage.

关 键 词： 水稻 蛋白质组 幼穗 双向电泳 质谱分析

领　　域： [农业科学]