| 转红色荧光蛋白基因唐鱼外源基因拷贝数的测定 |
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| 引用本文： | 樊佳佳,白俊杰,马冬梅,简清.转红色荧光蛋白基因唐鱼外源基因拷贝数的测定[J].生物技术通讯,2011,22(1):61-65. |
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| 作者姓名： | 樊佳佳 白俊杰 马冬梅 简清 |
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| 作者单位： | 中国水产科学研究院,珠江水产研究所,热带亚热带鱼类选育与养殖重点开放实验室,广东,广州,510380 |
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| 基金项目： | 国家高技术研究发展计划(2009AA10Z105) |
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| 摘 要： | 目的:测定转红色荧光蛋白基因唐鱼的外源基因拷贝数。方法:以杂合F5代和杂合F6代转红色荧光蛋白基因唐鱼为材料,以其外源插入片段(pDsRed-mylz2)与基因组插入位点5′侧翼区之间的边界序列作为特异性内参片段,同时以外源整合的红色荧光表达载体序列(pDsRed-mylz2)作为目的基因,采用实时荧光定量PCR技术测定外源基因整合的拷贝数。结果:运用外源基因与特异内参在同批次实时荧光定量PCR中的初始拷贝数比值,得到杂合F5代和杂合F6代转红色荧光蛋白基因唐鱼中插入的外源红色荧光表达载体序列pDsRed-mylz2的拷贝数均值均为3。结论:转红色荧光蛋白基因唐鱼的外源基因拷贝数为3。
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| 关 键 词： | 唐鱼 实时荧光定量PCR 外源基因 拷贝数 |
Estimating the Copy Number of Exogenous Gene in Transgenic Tanichthys albonubes Expressing the Red Fluorescent Protein |
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| FAN Jia-Jia,BAI Jun-Jie,MA Dong-Mei,JIAN Qing.Estimating the Copy Number of Exogenous Gene in Transgenic Tanichthys albonubes Expressing the Red Fluorescent Protein[J].Letters in Biotechnology,2011,22(1):61-65. |
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| Authors: | FAN Jia-Jia BAI Jun-Jie MA Dong-Mei JIAN Qing |
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| Institution: | FAN Jia-Jia,BAI Jun-Jie,MA Dong-Mei,JIAN Qing Key Laboratory of Tropical and Subtropical Fish Breeding & Cultivation,Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510380,China |
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| Abstract: | Objective:To estimate the copy number of exogenous gene in transgenic Tanichthys albonubes expressing the red fluorescent protein.Methods:The F5 and F6 generation heterozygous transgenic T.albonubes expressing the red fluorescent protein were used to estimate the copy number of exogenous gene by quantitative real-time PCR.The event-specific sequence was the border-sequence of pDsRed-mylz2 and 5' flanking region sequence,the objective gene was pDsRed-mylz2.Results:The F5 and F6 generation heterozygous transg... |
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| Keywords: | Tanichthys albonubes quantitative real-time PCR exogenous gene copy number |
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