| 山羊卵巢颗粒细胞差异表达基因消减文库的建立 |
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| 引用本文： | 庞训胜,王子玉,应诗家,张艳丽,闫益波,吴勇聪,孟立,王锋.山羊卵巢颗粒细胞差异表达基因消减文库的建立[J].南京农业大学学报,2010,33(6). |
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| 作者姓名： | 庞训胜 王子玉 应诗家 张艳丽 闫益波 吴勇聪 孟立 王锋 |
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| 作者单位： | 1. 南京农业大学羊业科学研究所,江苏南京210095;安徽科技学院动物科学学院,安徽,凤阳233100
2. 南京农业大学羊业科学研究所,江苏南京,210095 |
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| 基金项目： | 国家科技支撑计划项目(2008BADB2B04,2008BADB2B09); 江苏省高技术研究项目(BG2007324) |
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| 摘 要： | 为了从单卵泡水平建立非闭锁大小卵泡颗粒细胞基因表达的消减文库,选择繁殖性能正常、空怀高繁殖力黄淮山羊2只,以氯前列烯醇处理40h后取卵巢,计数卵泡并测量卵泡直径,钝性分离采集卵泡颗粒细胞,置于液氮内保存,取样后的卵泡部分组织采用原位缺口末端标记法(TUNEL)进行闭锁鉴定。选择直径6和4mm非闭锁卵泡,提取颗粒细胞mRNA,扩增cDNA,监测抑制消减杂交及其文库构建的试验环节。结果表明:消减文库的阳性克隆测序成功率100%,从正向文库中随机挑取96个克隆进行差异表达的筛选,发现12个全新表达序列标签;应用RT-PCR对已知序列的抑制素βA亚基基因(INHBA)和谷胱甘肽S-转移酶A1基因(GSTA1)进行表达水平测定,证实了颗粒细胞2个基因mRNA的表达模式与差异显示的结果相同。表明消减文库构建成功。
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| 关 键 词： | 山羊 卵巢 颗粒细胞 基因表达 消减文库 |
Construction of a subtractive library of genes differentially expressed in follicular granulosa cells of goat ovary |
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| PANG Xun-sheng,WANG Zi-yu,YING Shi-jia,ZHANG Yan-li,YAN Yi-bo,WU Yong-cong,MENG Li,WANG Feng.Construction of a subtractive library of genes differentially expressed in follicular granulosa cells of goat ovary[J].Journal of Nanjing Agricultural University,2010,33(6). |
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| Authors: | PANG Xun-sheng WANG Zi-yu YING Shi-jia ZHANG Yan-li YAN Yi-bo WU Yong-cong MENG Li WANG Feng |
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| Institution: | PANG Xun-sheng1,2,WANG Zi-yu1,YING Shi-jia1,ZHANG Yan-li1,YAN Yi-bo1,WU Yong-cong1,MENG Li1,WANG Feng1 (1. Institute of Sheep and Goats Science,Nanjing Agricultural University,Nanjing 210095,China,2. College of Animal Science,Anhui Science and Technology University,Fengyang 233100,China) |
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| Abstract: | In order to construct a subtractive cDNA library of genes differentially expressed in follicular granulosa cells between a single large and small unatresia follicles of goat ovary,two Huanghuai goats with normally reproductive ability and high prolificacy during estrous cycle were selected,of which the ovaries were taken out after treating with prostaglandin 40 hours. Follicles were counted and measured,then granulosa cells of antral follicles were collected by blunt dissection and stored in liquid nitrogen... |
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| Keywords: | goat ovary granulosa cell gene expression subtractive library |
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