| 表达新城疫病毒F基因的重组鸡痘病毒实时荧光定量PCR检测方法的建立 |
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| 引用本文： | 任静强,朱战波,贾鹏,赵权,袭莹,刘存霞,龙川,杜寿文,岳云强,鲁会军,金宁一.表达新城疫病毒F基因的重组鸡痘病毒实时荧光定量PCR检测方法的建立[J].中国兽医科学,2011(3). |
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| 作者姓名： | 任静强 朱战波 贾鹏 赵权 袭莹 刘存霞 龙川 杜寿文 岳云强 鲁会军 金宁一 |
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| 作者单位： | 吉林农业大学动物科技学院;军事医学科学院军事兽医研究所;黑龙江八一农垦大学动物科技学院; |
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| 基金项目： | 国家高技术研究发展计划(863)项目(2006AA10A205) |
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| 摘 要： | 采用荧光染料SYBR Green渗入法,通过对real-time PCR反应条件进行优化,建立了real-timePCR检测方法,用该方法定量分析新城疫病毒(NDV)F基因在重组鸡痘病毒(rFPV-F-VP0)第1、5、10、15、20代次中的表达整合情况。结果表明,建立的标准曲线呈现良好的重复性和特异性,与模板浓度呈现良好的线性关系。在线性浓度范围内,随着模板量的减少,其对应的Ct值相应增大,0.99<相关系数(r2)<1,0.8<扩增效率(E)<1.2,熔解曲线为单一特征峰型。本试验精确定量了外源基因在重组鸡痘病毒中的表达水平,为阐明重组鸡痘病毒的表达机理提供了理论基础。
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| 关 键 词： | 新城疫病毒 F基因 SYBR Green 实时荧光定量聚合酶链反应 鸡痘病毒 基因表达 |
Development of real-time PCR method for detection of recombinant fowlpox virus coexpressing F gene of Newcastle disease virus |
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| REN Jing-qiang,,ZHU Zhan-bo,JIA Peng,ZHAO Quan,XI Ying,LIU Cun-xia,LONG Chuan,DU Shou-wen,YUE Yun-qiang,LU Hui-jun,JIN Ning-yi.Development of real-time PCR method for detection of recombinant fowlpox virus coexpressing F gene of Newcastle disease virus[J].Veterinary Science in China,2011(3). |
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| Authors: | REN Jing-qiang ZHU Zhan-bo JIA Peng ZHAO Quan XI Ying LIU Cun-xia LONG Chuan DU Shou-wen YUE Yun-qiang LU Hui-jun JIN Ning-yi |
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| Institution: | REN Jing-qiang1,2,3,ZHU Zhan-bo2,JIA Peng2,ZHAO Quan1,XI Ying2,LIU Cun-xia2,LONG Chuan1,DU Shou-wen2,YUE Yun-qiang1,LU Hui-jun2,JIN Ning-yi2(1.College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China,2.Military Veterinary Institute,Academy of Military Medical Science,Changchun 130122,3.College of Animal Science and Technology,Heilongjiang August-First Land Reclamation University,Daqing 163319,China) |
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| Abstract: | In order to quantitatively analyze the expression of F gene of Newcastle disease virus(NDV) in recombinant fowlpox virus rFPV-F-VP0,the method with good reproducibility,high specificity and sensitivity standard was established after the real-time PCR reaction conditions were optimized by fluorescent dying methods of SYBR Green.The expression level of NDV F gene in the recombinant fowlpox virus rFPV-F-VP0 was quantitatively analyzed in passages 1,5,10,15 and 20,respectively.The results demonstrated that the ... |
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| Keywords: | Newcastle disease virus F gene SYBR Green real-time PCR fowlpox virus gene expression |
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