学术
小鼠单个GV期卵母细胞中ATP8基因的表达分析
洪灯[1] 宋绪华[2] 符史干[3] 景雅[4]
[1]海南医学院组胚学与胚胎学教研室,海口571101 [2]海南医学院附属医院眼科,海口571101 [3]海南医学院生理学教研室,海口571101 [4]山西医科大学组胚学与胚胎学教研室,太原030001
摘 要:
目的：探讨小鼠GV期卵母细胞线粒体中ATP8（ATP舍酶亚基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EcoR Ⅰ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。[著者文摘]
文章出处:
《现代生物医学进展》-2007年7卷10期 -1488-1490页
栏目信息:
文献标识码:
A
文章编号:
1673-6273（2007）10-1488-03
Analysis on Expression of ATP8 Gene in Single Oocyte in Mouse GV Stage
HONG Deng, SONG Xu-hua, FU Shi-gan, JING Ya （1. Department of Histology and Embryology, Hainan Medical College, Haikou 571101, China; 2. Department of Ophthalmology, Affiliated Hospital of Hainan Medical College, Haikou 570102, China; 3.Department of Physiology, Hainan Medical College, Haikou 571101, China, China; 4. Department of Histology and Embryology, Shanxi Medical University, Taiyuan, Shanxi 030001, China）
Abstract:
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.[著者文摘]
Key words:
Mouse; Germinal vesicle; Oocytes; ATPase subunit 8
基金资助:
国家自然科学基金项目（No.30360039）致谢：衷心感谢海南医学院转基因实验室董克家研究员给予理论和技术上的指导.海南医学院媛老师、符皎荣老师、崔玉宝老师、郑小桃老师、钟南田老师、吴亚强同学和王景景同学的支持和帮助.
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