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Comparative proteomic profiling during ovarian development of the shrimp Metapenaeus ensis

机译:虾对虾对虾卵巢发育过程中的蛋白质组比较分析

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Two-dimensional electrophoresis and mass spectrometry were used to identify proteins that are differentially expressed during ovarian maturation in Metapenaeus ensis. 87 spots with consistently significant quantitative differences (a parts per thousand yen1.5-fold for vol%) among stage I, III and V ovaries were chosen for MS/MS analysis. 45 spots were significantly matched to known proteins in the database (Mascot score > 40). Half of them were down-regulated, in contrast to 9 out of 45 proteins that were up-regulated as ovarian maturation proceeded. Functionally, these identified proteins could be classified into five major groups, including cytoskeleton (11 %), metabolism (18 %), signal transduction (32 %), gene expression (14 %) and immune response (7 %). Among the differentially expressed reproduction-related proteins, the mRNA expression level of cellular retinoic acid/retinol binding protein in M. ensis (MeCRABP) during ovarian maturation was further characterized by quantitative real-time PCR. It was down-regulated during ovarian maturation. In situ hybridization further revealed that MeCRABP transcript was localized in ooplasm of previtellogenic oocytes but not in vitellogenic oocytes. These results demonstrate the application of proteomic analysis for identification of proteins involved in shrimp ovarian maturation and they provide new insights into ovarian development.RI Chu, Ka Hou/B-8010-2011; WU, Longtao/G-6477-2014OI WU, Longtao/0000-0003-3463-4127
机译:二维电泳和质谱法用于鉴定Metapenaeus ensis在卵巢成熟过程中差异表达的蛋白质。在I,III和V期卵巢之间选择了87个点,这些点的定量差异始终是显着的(千分之1.5的体积百分比),用于MS / MS分析。 45个斑点与数据库中的已知蛋白质显着匹配(Mascot得分> 40)。与卵巢成熟过程中上调的45种蛋白质中的9种相比,其中的一半被下调。从功能上讲,这些鉴定出的蛋白质可分为五个主要类别,包括细胞骨架(11%),新陈代谢(18%),信号转导(32%),基因表达(14%)和免疫应答(7%)。在差异表达的生殖相关蛋白中,通过实时荧光定量PCR进一步鉴定了卵巢成熟过程中中华绒螯蟹中细胞视黄酸/视黄醇结合蛋白的mRNA表达水平。在卵巢成熟过程中它被下调。原位杂交进一步揭示,MeCRABP转录物位于玻璃体形成前卵母细胞的卵质中,而不存在于卵黄形成卵母细胞中。这些结果证明了蛋白质组学分析在鉴定与虾卵巢成熟有关的蛋白质中的应用,它们为卵巢的发育提供了新的见识。RI Chu,Ka Hou / B-8010-2011; WU,Longtao / G-6477-2014OI WU,Longtao / 0000-0003-3463-4127

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