A novel method for constructing murine cDNA library enriched with maternal mRNAs exhibiting de novo independent post-fertilization polyadenylation.
Recently, mouse maternal mRNAs such as SSEC-D, Spin, beta-catenin, Ptp4a1, 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.
Sakurai
Takayuki
T
Division of Basic Molecular Science and Molecular Medicine, School of Medicine, Tokai University, Bohseidai, Isehara, Kanagawa 259-1193, Japan. sakurai@is.icc.u-tokai.ac.jp
Sato
Masahiro
M
Kimura
Minoru
M
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D018076:DNA, Complementary / Q000235:genetics* / Q000378:metabolism; D005306:Fertilization / Q000235:genetics*; D015723:Gene Library; D051379:Mice; D009865:Oocytes / Q000378:metabolism; D010063:Ovum / Q000378:metabolism; D026723:Polyadenylation / Q000235:genetics* / Q000502:physiology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism
D018076:DNA, Complementary; D012333:RNA, Messenger