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通过消减免疫揭示非洲爪蟾中的局部母体蛋白。

Localized maternal proteins in Xenopus revealed by subtractive immunization.

作者信息

Denegre J M, Ludwig E R, Mowry K L

机构信息

Department of Molecular Biology, Cell Biology & Biochemistry, Brown University, Providence, Rhode Island 02912, USA.

出版信息

Dev Biol. 1997 Dec 15;192(2):446-54. doi: 10.1006/dbio.1997.8773.

Abstract

It has long been appreciated that the localization of cytoplasmic determinants in the egg can provide the foundation for patterning in the embryo. Differences in cell fate among the early blastomeres are thus a consequence of asymmetric distributions of informational molecules prior to fertilization. The frog egg has a single axis of asymmetry present prior to fertilization, the animal/vegetal axis, and the localization of developmental information appears to be polarized along this axis. Such developmental information can be localized as either RNA or protein; localized RNAs are well documented in the Xenopus oocyte, and some are thought to play roles in axial patterning. While it is apparent that not all of the localized maternal components are RNAs, much less is known about maternal proteins that might be localized in the egg. In the present study, we have taken a novel approach to identify localized maternal proteins within the Xenopus egg. Using a subtractive immunization strategy, we have generated monoclonal antibodies which recognize antigens that are restricted to the vegetal cortex of fertilized eggs. Analysis of biogenesis during oogenesis reveals two distinct patterns of localization to the cortex. At least three of these localized antigens are proteins, and these localized proteins could represent maternal determinants with roles in patterning.

摘要

长期以来,人们一直认识到卵子中细胞质决定因子的定位可为胚胎模式形成提供基础。因此,早期卵裂球之间细胞命运的差异是受精前信息分子不对称分布的结果。蛙卵在受精前有一个单一的不对称轴,即动物/植物轴,发育信息的定位似乎沿此轴呈极化分布。这种发育信息可以定位为RNA或蛋白质;在非洲爪蟾卵母细胞中,定位RNA已有充分记载,并且一些RNA被认为在轴模式形成中发挥作用。虽然显然并非所有定位的母体成分都是RNA,但对于可能定位在卵子中的母体蛋白质却知之甚少。在本研究中,我们采用了一种新方法来鉴定非洲爪蟾卵内定位的母体蛋白质。利用消减免疫策略,我们产生了单克隆抗体,这些抗体识别仅限于受精卵植物皮层的抗原。对卵子发生过程中生物合成的分析揭示了两种不同的皮层定位模式。这些定位抗原中至少有三种是蛋白质,并且这些定位蛋白质可能代表在模式形成中起作用的母体决定因子。

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