| 竞争RT-PCR方法检测奶牛肝载脂蛋白E基因mRNA变化的内参构建 |
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| 引用本文： | 王雪莹,孙玉成,陈仕均,王哲,杨连玉.竞争RT-PCR方法检测奶牛肝载脂蛋白E基因mRNA变化的内参构建[J].中国兽医杂志,2006,42(12):5-7. |
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| 作者姓名： | 王雪莹 孙玉成 陈仕均 王哲 杨连玉 |
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| 作者单位： | 吉林大学农学部畜牧兽医学院,吉林,长春,130062 |
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| 摘 要： | 构建定量逆转录-聚合酶链反应（RT—PCR）的内参标准，用以检测围产期奶牛肝脂代谢过程中ApoE基因mRNA表达的变化情况。应用限制性内切酶ApaI消化重组质粒PMD—ApoE757使其缺失3个小片段。然后再回收较大的片段和酶切后的PMD—ApoE757线性片段，回收的两个片段用T4连接酶连接后，成功构建了PMD—ApoE487重组质粒。
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| 关 键 词： | ApoE基因 竞争模板 定量RT-PCR 克隆 |
| 文章编号： | 0529-6005（2006）12-0005-03 |
| 收稿时间： | 07 12 2005 12:00AM |
| 修稿时间： | 2005年7月12日 |
An internal reference technique for quantifying specific mRNA by RT-PCR with application to the lipoprotein E gene |
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| WANG Xue-ying,SUN Yu-cheng,CHEN Shi-jun,WANG Zhe,YANG Lian-yu.An internal reference technique for quantifying specific mRNA by RT-PCR with application to the lipoprotein E gene[J].Chinese Journal of Veterinary Medicine,2006,42(12):5-7. |
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| Authors: | WANG Xue-ying SUN Yu-cheng CHEN Shi-jun WANG Zhe YANG Lian-yu |
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| Institution: | College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China |
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| Abstract: | The objective of the present study is to construct an internal standard which can be used as a competitor for ApoE. Three small fragments were cut from a 757 bp ApoE recombinant plasmid by restriction enzyme ApaI. Then, the larger linear fragment and liner 757bp ApoE fragment was ligated by T4DNA ligase. Thus we construct PMD-ApoE487 recombinant plasmid successfully. It can be applied for quantification of ApoE mRNA in dairy cows. |
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| Keywords: | ApoE inter-reference quantitive RT-PCR clone |
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