Temeles G L, Ram P T, Rothstein J L, Schultz R M
Department of Biology, University of Pennsylvania, Philadelphia 19104-6018.
Mol Reprod Dev. 1994 Feb;37(2):121-9. doi: 10.1002/mrd.1080370202.
Little is known about the repertoire of genes expressed following zygotic gene activation, which occurs during the two-cell stage in the mouse. As an initial attempt to isolate novel genes, we used previously prepared two-cell and two-cell subtraction cDNA libraries (Rothstein et al., Genes Dev 6:1190-1201, 1992) to isolate a panel of seven cDNA clones. Three cDNAs had no match in the current DNA sequence data banks and three others revealed sequence homology to portions of sequences in the data banks. One cDNA was 90% homologous to the ras-related gene Krev/rap 1A. The temporal patterns of expression of these genes during oocyte maturation and preimplantation development were analyzed by a reverse transcription-polymerase chain reaction (RT-PCR) assay developed to measure relative levels of mRNAs. Three distinct temporal patterns of expression, designated Classes 1-3, were found. The two Class 1 genes displayed an actin-like pattern, with a gradual decline in expression during oocyte maturation and through the two-cell stage, followed by increases at the eight-cell and/or blastocyst stages. The four genes in Class 2 were expressed at relatively high levels during oocyte maturation and through the one-cell stage and then declined abruptly between the one- and two-cell stages; an increase then occurred at the eight-cell and/or blastocyst stages. The expression of the gene in Class 3 declined during oocyte maturation, but then showed a transient increase at the one-cell stage, with only a very slight increase in synthesis at either the eight-cell or blastocyst stage.
关于合子基因激活后表达的基因库知之甚少,合子基因激活发生在小鼠的二细胞阶段。作为分离新基因的初步尝试,我们使用先前制备的二细胞和二细胞消减cDNA文库(罗斯坦等人,《基因与发育》6:1190 - 1201,1992)分离出一组七个cDNA克隆。三个cDNA在当前的DNA序列数据库中没有匹配项,另外三个显示出与数据库中部分序列的同源性。一个cDNA与ras相关基因Krev/rap 1A有90%的同源性。通过一种用于测量mRNA相对水平的逆转录 - 聚合酶链反应(RT-PCR)分析方法,分析了这些基因在卵母细胞成熟和植入前发育过程中的表达时间模式。发现了三种不同的表达时间模式,分别命名为1 - 3类。两个1类基因呈现出肌动蛋白样模式,在卵母细胞成熟过程中以及通过二细胞阶段时表达逐渐下降,随后在八细胞和/或囊胚阶段增加。2类中的四个基因在卵母细胞成熟过程中以及通过单细胞阶段时表达水平相对较高,然后在单细胞和二细胞阶段之间突然下降;随后在八细胞和/或囊胚阶段出现增加。3类基因的表达在卵母细胞成熟过程中下降,但在单细胞阶段出现短暂增加,在八细胞或囊胚阶段合成仅略有增加。