| 树鼩IL-2TaqMan探针实时荧光定量RT-PCR检测方法的建立 |
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| 引用本文： | 黄晓燕,徐娟,王文广,殷安国,李晓飞,孙晓梅,夏雪山,代解杰. 树鼩IL-2TaqMan探针实时荧光定量RT-PCR检测方法的建立[J]. 中国比较医学杂志, 2013, 0(7): 36-40,49 |
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| 作者姓名： | 黄晓燕 徐娟 王文广 殷安国 李晓飞 孙晓梅 夏雪山 代解杰 |
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| 作者单位： | 中国医学科学院/北京协和医学院医学生物学研究所,云南省重大传染病疫苗研发重点实验室,树鼩种子资源中心;昆明理工大学生命科学与技术学院 |
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| 基金项目： | 国家科技支撑计划项目(编号:2009BAI83B02-21;2011BAI15B01-21;2012BAI39B01);云南省科技计划重点项目(编号:2006PT07-2);云南省应用基础面上项目(编号:2011FZ211) |
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| 摘 要： | 目的 建立检测树鼩IL-2基因TaqMan探针实时荧光定量PCR方法.方法 以经ConA诱导培养的树鼩淋巴细胞总RNA为材料,设计特异性引物,通过RT-PCR技术克隆出树鼩IL-2基因,以构建的树鼩IL-2基因质粒为标准品,建立标准曲线,并进行灵敏度检测.结果 建立了树鼩IL-2基因mRNA表达实时荧光定量PCR检测方法,此法检测灵敏度高,线性范围可达102～109拷贝/ul.结论 成功建立了树鼩IL-2实时荧光定量PCR方法,此方法灵敏度高、特异性强,检测周期短,为探讨IL-2作用的分子作用机制奠定基础.
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| 关 键 词： | 树鼩 IL-2 实时荧光定量 PCR |
Establishment of a real-time flurescent quantitative RT-PCR assay for detection of tree threw IL-2 gene |
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| Huang Xiao-yan,Xu Juan,Wang Wen-guang,Yin An-guo,Li Xiao-fei,Sun Xiao-mei,Xia Xue-shan,Dai Jie-jie. Establishment of a real-time flurescent quantitative RT-PCR assay for detection of tree threw IL-2 gene[J]. Chinese Journal of Comparative Medicine, 2013, 0(7): 36-40,49 |
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| Authors: | Huang Xiao-yan Xu Juan Wang Wen-guang Yin An-guo Li Xiao-fei Sun Xiao-mei Xia Xue-shan Dai Jie-jie |
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| Affiliation: | 1(1.Center of Tree Shrew Germplasm Resources,Institute of Medical Biology,the Chinese Academy of Medical Science and Peking Union Medical College.Yunnan Key Laboratory of Vaccine Research and Development on Severe Infectious Diseases.Kunming 6501182.Faculty of Life Science and Technology,Kunming University of Science and Technology,Kunming 650500,China) |
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| Abstract: | Objective To establish a real-time fluorescent quantitative RT-PCR assay to detect tree threw （tupaiabelangeri） IL-2. Methods Total RNA was isolated from Con A-stimulated tree shrews＇ spleen lymphocytes. The conservative encoding sequence of interleukin-2 （IL-2） was amplified by RT-PCR, and successfully cloned into pMD-19T vector. The established IL-2 gene vector was used as standard substance and the standard curves were established for the sensitivity evaluation. Results A real-time fluorescence quantitative method of fast, sensitive and reliable detection system was set up. The sensitivity of this method for creating IL-2 gene of tree shrew was 102 ~ 109copies. Conclusions A real- time fluorescence quantitative method of tree shrew IL-2 was successfully established. This sensitive method can serve as a molecular foundation for the study of IL-2 and its role in many clinical diseases. |
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| Keywords: | Tree shrew IL-2 Real-time flurescent quantitative RT-PCR assay |
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