| 黄瓜CMV复制酶基因相对表达定量PCR方法的建立 |
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| 引用本文： | 王军辉,张桂华,杜胜利,王永琦,张显.黄瓜CMV复制酶基因相对表达定量PCR方法的建立[J].中国瓜菜,2010,23(3):15-18. |
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| 作者姓名： | 王军辉 张桂华 杜胜利 王永琦 张显 |
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| 作者单位： | 1. 西北农林科技大学园艺学院,陕西杨凌,712100
2. 天津科润黄瓜研究所,天津,300192 |
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| 基金项目： | 天津市应用基础研究计划项目，国家"863"计划项目 |
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| 摘 要： | 为了对黄瓜感染CMV植株体内CMV复制酶基因表达水平进行测定,建立CMV复制酶基因的相对表达定量检测技术,并用该方法开展黄瓜植株CMV抗性鉴定试验,以18SrRNA基因为对照,CMV复制酶基因为目标基因,设计引物,探索SYBRGreen实时荧光定量PCR反应体系,同时对CMV复制酶基因进行均一化处理,利用荧光阈值(Q值)计算p-防御素基因的相对表达量。结果表明:利用SYBR GreenⅠ实时荧光定量PCR技术测定黄瓜CMV复制酶基因相对表达定量具有较高的准确性,耗时短,该技术可应用于CMV抗性植株的筛选。
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| 关 键 词： | 实时荧光定量PCR CMV复制酶基因 相对表达量 黄瓜 |
Detection of Relative Expression of Cucumber Mosaic Virus Replicase Gene by SYBR Green I RT-PCR |
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| WANG Jun-hui,ZHANG Gui-hua,DU Sheng-li,WANG Yong-qi,ZHANG Xian.Detection of Relative Expression of Cucumber Mosaic Virus Replicase Gene by SYBR Green I RT-PCR[J].China Cucurbits And Vegetables,2010,23(3):15-18. |
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| Authors: | WANG Jun-hui ZHANG Gui-hua DU Sheng-li WANG Yong-qi ZHANG Xian |
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| Institution: | WANG Jun-hui1,ZHANG Gui-hua2,DU Sheng-li2,WANG Yong-qi1,ZHANG Xian1(1.College of Horticulture,Northwest Science , Technology University of Agriculture , Forestry,Yangling,Shaanxi,712100,China,2.Tianjin Kernel Cucumber Research Institute,Tianjin,300192,China) |
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| Abstract: | To detect the relative expression of Cucumber mosaic virus(CMV)replicase gene in cucumber,a method based on real time PCR with SYBR Green I was developed.Two primer sets were designed for 18S and CMV and relative standard curve was figured in order to quantify the express of CMV replicase of 189 samples.The results indicated that the SYBR GreenⅠ RT-PCR is stable and reliable method for detecting the relative expression of CMV replicase gene,and the method can be used to screen the CMV resistant cucumber plants. |
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| Keywords: | Real-time quantitative PCR CMV Relative expression Cucumber |
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