| 西农萨能奶山羊泌乳中期和后期乳腺组织差异基因的筛选 |
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| 引用本文： | 武会娟,罗军,赵旺生,王伟.西农萨能奶山羊泌乳中期和后期乳腺组织差异基因的筛选[J].畜牧兽医学报,2010,41(1). |
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| 作者姓名： | 武会娟 罗军 赵旺生 王伟 |
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| 作者单位： | 1. 西北农林科技大学动物科技学院,杨凌,712100;北京市理化分析测试中心,北京,100193
2. 西北农林科技大学动物科技学院,杨凌,712100 |
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| 基金项目： | 国家863项目"奶山羊乳腺脂肪酸代谢基因网络分析"，国家科技支撑计划项目，北京市科学技术研究院创新团队计划 |
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| 摘 要： | 旨在利用抑制性削减杂交(SSH)技术筛选西农萨能奶山羊泌乳中期和后期的差异表达基因,并用实时定量PCR(Q-PCR)分析差异基因的表达丰度,探讨奶山羊不同泌乳阶段乳腺组织的基因表达规律。本研究以西农萨能奶山羊泌乳中期和后期乳腺组织的mRNA互为检测子(Tester)和驱动子(Driver)构建cDNA消减文库(M-L和L-M),随机挑选克隆测序,进行序列比对分析,并检测部分差异基因在乳腺组织中的表达丰度。结果,成功构建了泌乳中期和后期的cDNA文库,随机挑选30个克隆测序,得到M-L和L-M文库中与细胞凋亡、抗氧化、脂类代谢、能量代谢等生理过程相关的差异基因,对其中的6个基因进行实时定量分析,发现5个均为阳性克隆,表达水平增加了1.3~5.5倍不等。结果表明,利用SSH技术成功构建了泌乳中期和后期乳腺组织正反向消减cDNA文库,筛选出24个差异基因,为进一步研究奶山羊乳腺组织基因调控机理奠定了基础。
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| 关 键 词： | 西农萨能羊 乳腺组织 抑制性削减杂交 差异表达基因 实时定量PCR |
Screening of Differentially Expressed Genes in Mammary Gland of Xinong Saanen Dairy Goat at Middle and Late Lactation Stages |
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| WU Hui-juan,LUO Jun,ZHAO Wang-sheng,WANG Wei.Screening of Differentially Expressed Genes in Mammary Gland of Xinong Saanen Dairy Goat at Middle and Late Lactation Stages[J].Acta Veterinaria et Zootechnica Sinica,2010,41(1). |
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| Authors: | WU Hui-juan LUO Jun ZHAO Wang-sheng WANG Wei |
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| Institution: | WU Hui-juan1,2,LUO Jun1,ZHAO Wang-sheng1,WANG Wei1(1.College of Animal Science , Technology,Northwest A&F University,Yangling 712100,China,2.Beijing Center for Physical , Chemical Analysis,Beijing 100193,China) |
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| Abstract: | The objective of this study was to use suppression subtractive hybridization(SSH) to screen the differentially expressed genes in mammary gland of Xinong Saanen Dairy goats at the middle and late lactation stages,and the expression abundance of several genes were analyzed by real-time quantitative PCR(Q-PCR) to explore the gene expression mechanism of mammary gland in the different lactation stages of dairy goats.The subtracted cDNA libraries(M-L and L-M) were constructed by using mRNA of mammary gland of X... |
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| Keywords: | Xinong Saanen Dairy goat mammary gland suppression subtractive hybridization differentially expressed gene real-time quantitative PCR |
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