Analysis of gene expression in the preimplantation mouse embryo: use of mRNA differential display.
Analysis of gene expression in the preimplantation mouse embryo: use of mRNA differential display.
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DOI：
10.1073/pnas.91.12.5456
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发表时间：
1994-06
影响因子：
11.1
通讯作者：
J. Zimmermann;R. Schultz
中科院分区：
文献类型：
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作者：
J. Zimmermann;R. Schultz
生物材料的缺乏阻碍了植入前胚胎发生过程中差异基因表达的分析。我们报告了对最近描述的 mRNA 差异显示方法（Liang, P. & Pardee, A. B. (1992) Science 257, 967-971）的修改，以分析小鼠植入前发育过程中的差异基因表达。该方法检测扩增子表达时间模式的适当变化，通过 DNA 序列分析，该扩增子是细胞角蛋白内切 A，该基因的表达时间模式先前已通过 S1 核酸酶消化确定。此外，该方法还鉴定了可能代表以下基因的扩增子：(i) 编码母体 mRNA，(ii) 早期和晚期合子基因激活的产物，(iii) 其表达在八细胞阶段最大（即，以阶段特异性方式表达），以及 (iv) 其表达在囊胚中最大。除了内切 A 之外，对这些扩增子的序列分析表明，显示与其母体 mRNA 一致的表达时间模式的扩增子是线粒体 F1 ATP 合酶的 α 亚基。
The analysis of differential gene expression during preimplantation embryogenesis has been hindered by the paucity of biological material. We report modifications of the recently described mRNA differential display method (Liang, P. & Pardee, A. B. (1992) Science 257, 967-971) to analyze differential gene expression during mouse preimplantation development. The method detects the appropriate changes in the temporal pattern of expression of an amplicon that by DNA sequence analysis is the cytokeratin endo A, a gene whose temporal pattern of expression has been previously determined by S1 nuclease digestion. In addition, this method identifies amplicons that likely represent genes (i) that encode maternal mRNAs, (ii) that are products of early and late zygotic gene activation, (iii) whose expression is greatest during the eight-cell stage (i.e., expressed in a stage-specific manner), and (iv) whose expression is greatest in the blastocyst. In addition to endo A, sequence analysis of these amplicons reveals that an amplicon that displays a temporal pattern of expression consistent with it being a maternal mRNA is the alpha subunit of the mitochondrial F1 ATP synthase.