摘要
目的:探讨小鼠GV期卵母细胞线粒体中ATP8(ATP合酶亚基8)基因的表达情况。方法:应用挤压法从卵巢中分离获得生发泡期(germinal vesicle,GV)卵母细胞;用RT-PCR检测GV期单个卵母细胞中ATP8基因的表达:其中cDNA的合成分两种方法进行:一是将GV期单个卵母细胞直接进行RT合成cDNA,二是先用DNA酶加EcoRⅠ酶祛除mtDNA和核DNA后再进行RT;回收产物构建克隆质粒并测序。结果:1.5%琼脂糖电泳显示、测序结果均表明ATP8基因在GV期卵母细胞中有表达。结论:小鼠GV期卵母细胞特异表达的ATP8基因可能与卵母细胞的正常发育成熟相关。
Objective： To investigate the expression of the ATP8 gene in the mitochondria in the single oocyte in mouse germinal vesicle GV stage. Methods： Oocytes in the GV stage were separated from ovary by squeezing method.In mouse germinal vesicle GV stage, the expression of ATP8 gene in the mitochondria in the single oocyte was detected by RT-PCR, in which, cDNA was synthesized with two methods： one was the single GV-stage oocyte directly to be placed RT, the other was to perform RT after eliminating mtDNA and nucleus DNA with the EcoRI enzyme and Dnase. And the product of RT-PCR was cloned and sequenced. Results： The results of 1.5% agarose gel electrophoresis and sequencing showed that ATP8 gene was expressed in GV-stage oocytes. Conclusion： The specific expression of ATP8 gene in GV-stage oocyte might be related to normal development maturation of oocyte.
出处
《现代生物医学进展》
CAS
2007年第10期1488-1490,共3页
Progress in Modern Biomedicine
基金
国家自然科学基金项目(No.30360039)
