【摘要】 目的:探讨小鼠卵母细胞发育成熟与18S rRNA基因表达的关系。方法:根据Genbank中公布的小鼠18S rRNA保守区序列（363bp）,用DNAclub软件设计两对特异性引物,RT-PCR检测单个GV、MⅠ期卵母细胞18S rRNA的表达;取单个MⅠ期卵母细胞二次PCR产物连接至pTG19-T载体、转化大肠杆菌感受态细胞,取阳性克隆进行测序。结果:RT-PCR检测显示18S rRNA基因在小鼠单个GV期、MⅠ期卵母细胞中均有表达,且在未成熟卵母细胞中,MⅠ期的表达明显强于GV期的表达。测序结果表明小鼠单个卵母细胞表达的18S rRNA基因与参考序列（Genbank NR₀03278）完全一致。结论:小鼠MⅠ期之后的卵母细胞发育需要蛋白质的合成,卵母细胞的成熟可能与核糖体18S基因表达有关。更多还原

【Abstract】 Objective:To determine whether 18S rRNA is expressed in single immature oocytes in germinal vesicle（GV） and metaphaseⅠ（MI）or not.Methods:According to the conservative sequence coding for 18S rRNA of mus musculus（363 bp）,two pairs of primers used in RT-PCR were designed by DNAdub software.Subsequently,the expression of 18S rRNA was detected in single immature oocytes in GV and MⅠstage by RT-PCR.The second-PCR products from single immature oocytes in MⅠstage were inserted into pTG19-T vector,transformed into E.coil,and the positive recombinant plasmids were screened and sequenced,Results:The gene fragment encoding for 18S rRNA presented both in single GV and MⅠoocyte.But in immature oocytes,its expression in MⅠstage was higher than that in GV stage.The sequenced results were consistent with the conservative sequence of Genbank NR₀03278（791bp～1153bp）.Conclusion:The development of oocytes needs protein after metaphase stage,and its maturation may be related with 18S rRNA expression.更多还原