Analysis of gene expression in rabbit nuclear transfer embryos: Use of single-embryo mRNA differential display.
Lack of or abnormal expression of developmentally important genes is believed to hamper early development of the nuclear transfer (NT) embryo. To identify stage-specific genes in rabbit NT embryo development, mRNA differential display was used to compare the mRNA content of rabbit NT embryos at different developmental stages, from Metaphase II oocytes to 8-16-cell stage embryos. Thirty-four zygotic transcripts, which abruptly appeared at the 8-16-cell stage in rabbit NT embryos, were isolated; 11 of these were potential novel genes with no matches in the current databases. Of the remaining 23, 12 were matched with established sequence tags with functions uncharacterized and the other 11 were homologous to those in the European Molecular Biology Laboratory (EMBL) and GenBank databases. The differential expression of eight of the 34 amplicons were confirmed by reverse Northern blotting, and four positive clones were validated. Previous studies and present data indicated that these three genes were probably related to preimplantation rabbit embryo development.
Li
Wen Yong
WY
Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100080, China.
Yu
Wei Dong
WD
Qi
Bing
B
He
Xin
X
Wang
Yu Ge
YG
Liu
Gui Sheng
GS
Du
Miao
M
Chen
Qing Xuan
QX
D000818:Animals; D001483:Base Sequence; D001755:Blastocyst; D015152:Blotting, Northern; D002467:Cell Nucleus / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D019992:Databases as Topic; D004622:Embryo, Mammalian / Q000502:physiology*; D005314:Embryonic and Fetal Development; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D005821:Genetic Techniques; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D013997:Time Factors
D018076:DNA, Complementary; D012333:RNA, Messenger