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不同生理状态乳牛卵巢卵母细胞基因的差异表达
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张进隆1，2，赵兴绪1*，陶金忠1，杨永新1，张勇1
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（1.甘肃农业大学 动物医学院，甘肃 兰州730070；2.甘肃畜牧工程职业技术学院，甘肃 武威733006）
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摘要：
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为了研究不同生理状态下的卵巢卵母细胞基因的表达情况，采集乳牛的卵巢，分离了卵母细胞，以单个卵母细胞的mRNA作为模板，用设计的随机引物和锚定引物，采用一步法RT-PCR扩增了卵母细胞基因。经聚丙烯酰胺凝胶电泳检测，发现有功能黄体和无黄体卵巢卵母细胞基因的电泳条带之间无明显差异，对一条明显差异条带进行回收、纯化，将纯化的PCR产物连接到T载体,阳性克隆经鉴定后，进行测序和同源性比较。结果显示，与体外培养的牛早期胚胎的一条序列具有很高的同源性。 |
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关键词：
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乳牛；卵母细胞；基因表达 |
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中图分类号：Q 954.4：Q 786 文献标识码：
A 文章编号：1673-4696(2008)02-0151-04 |
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Differential expression of oocyte gene in dairy cows
at different physiological states |
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ZHANG Jin-long1,2,ZHAO Xing-xu1,TAO
Jin-zhong1,YANG Yong-xin1,ZHANG Yong1 |
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(1.College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China;
2.Gansu Polytechnical College of Animal Husbandry and Engineering，Wuwei 733006，China)
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Abstract: To detect gene expression of oocyte from ovaries in dairy cow at different physiological states,ovaries were collected from the slaughterhouse and divided into functional corpus luteum and non-corpus luteum.Oocytes were separated from the ovaries,using mRNA of single oocyte lysates as template,with Oligo(dt) and random primer,the oocyte genes were amplified successfully by one-step RT-PCR.The amplified products were detected with the polyacrylamide gel electrophoresis(PAGE) without obvious difference.After selection,recovery and purification,a differential PCR product was cloned into T-vector,sequenced and analyzed in homology.The result showed that the differentially expressed gene shared high identity with one sequence expressed by the early bovine embryo-in-vitro. |
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Key words:dairy cow;oocyte;gene expression |
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