| 反向PCR鉴定piggyBac介导的哺乳动物细胞转基因研究 |
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| 引用本文： | 郝碧芳,王猛.反向PCR鉴定piggyBac介导的哺乳动物细胞转基因研究[J].河南农业科学,2011,40(10):141-143,152. |
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| 作者姓名： | 郝碧芳 王猛 |
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| 作者单位： | 1. 江苏科技大学蚕业研究所,江苏镇江212018;中国农业科学院蚕业研究所,江苏镇江212018
2. 江苏科技大学蚕业研究所,江苏镇江,212018 |
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| 基金项目： | 江苏科技大学科技计划项目(35180905) |
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| 摘 要： | 利用反向PCR技术,研究piggyBac介导的转基因哺乳动物中外源基因整合位点信息.首先在转座子piggyBac中插入巨细胞病毒(CMV)启动子元件驱动的绿色荧光蛋白基因(EGFP),构建用于转染哺乳动物细胞的转基因载体pXL - CMV - EGFP；同时构建以CMV启动子驱动的转座酶基因载体.将2种载体以脂质体共转...
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| 关 键 词： | piggyBac 反向PCR 稳定表达 转基因 哺乳动物 |
Determination of Transgenic Research Mediated by piggyBac in Mammalian Cells by Reverse PCR |
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| HAO Bi-fang,WANG Meng.Determination of Transgenic Research Mediated by piggyBac in Mammalian Cells by Reverse PCR[J].Journal of Henan Agricultural Sciences,2011,40(10):141-143,152. |
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| Authors: | HAO Bi-fang WANG Meng |
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| Institution: | HAO Bi-fang1,2,WANG Meng1(1.Sericultural Research Institute,Jiangsu University of Science and Technology,Zhenjiang 212018,China,2.Sericultural Research Institute,Chinese Academy of Agricultural Sciences,China) |
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| Abstract: | To analyze the integration site information,reverse PCR was performed in the transgenic research.EGFP gene promoted by cytomegaoviyns virus(CMV) promoter was cloned into the insect transposon vector piggyBac to construct the transgenic vector pXL-CMV-EGFP.The transposase gene driven by CMV promoter was also constructed.Then they were co-transfected into the human embryonic kidney(HEK) 293 cells.Transfection of pXL-CMV-EGFP only was used as the negtive control and the fluoresences were observed in both treat... |
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| Keywords: | piggyBac Reverse PCR Stable expression Transgene Mammal |
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