A novel method for constructing murine cDNA library enriched with maternal mRNAs exhibiting de novo independent post-fertilization polyadenylation
A novel method for constructing murine cDNA library enriched with maternal mRNAs exhibiting de novo independent post-fertilization polyadenylation
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DOI：
10.1016/j.bbrc.2004.12.057
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发表时间：
2005-02-18
影响因子：
3.1
通讯作者：
Kimura, M
中科院分区：
文献类型：
--
作者：
Sakurai, T;Sato, M;Kimura, M
最近，发现小鼠母体 mRNA（例如 SSEC-D、Spin、β-catenin、Ptp4al 和 Maid）在受精后表现出从头独立的聚腺苷酸化。为了全面了解受精后多腺苷酸化事件，我们开发了一种构建小鼠受精卵 cDNA 文库的新方法，该文库富含表现出从头独立多腺苷酸化的 cDNA。作为一项初步研究，我们分离出至少四种新的母体 mRNA，这些 mRNA 在受精 1 细胞卵中表现出聚腺苷酸尾的延伸。此外，在此阶段观察到母体 RNA 的各种类型的聚腺苷酸化，表明存在调节母体 mRNA 聚腺苷酸尾长度的新机制。这是成功构建 cDNA 文库的第一份报告，该文库富含来自受精后小鼠卵子的新聚腺苷酸化的母体 mRNA。该 cDNA 文库可用于对哺乳动物母体 RNA 受精后多聚腺苷酸化的机制进行分子分析。 (C) 2004 Elsevier Inc. 保留所有权利。
Recently, mouse maternal mRNAs such as SSEC-D, Spin, beta-catenin, Ptp4al, and Maid have been found to exhibit de novo independent polyadenylation after fertilization. To obtain an overall picture of post-fertilization polyadenylation events, we developed a novel method for constructing murine fertilized egg cDNA library enriched with cDNAs exhibiting de novo independent polyadenylation. As a pilot study, we isolated at least four new maternal mRNAs exhibiting extension of poly(A) tail in fertilized 1-cell eggs. Moreover, various types of polyadenylation of maternal RNAs were observed at this stage, suggesting the presence of novel mechanisms for regulating the length of poly(A) tails of maternal mRNA. This is the first report of successful construction of a cDNA library enriched with newly polyadenylated maternal mRNAs derived from post-fertilized mouse eggs. This cDNA library will be useful for molecular analysis of the mechanisms underlying post-fertilization polyadenylation of mammalian maternal RNAs. (C) 2004 Elsevier Inc. All rights reserved.