| Production of transgenic mice carrying green fluorescence proteingene by a lentiviral vector-mediated approach |
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| 作者姓名： | ZHANG Jingzhi GUO Xinbing XIE Shuyang ZHU Yiwen HUANG Ying WANG Shu REN Zhaorui |
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| 作者单位： | Shanghai Institute of Medical Genetics, Shanghai JiaoTong University, Shanghai 200040, China |
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| 摘 要： | A pseudo-lentivirus, which carries green fluorescence protein (GFP) expressing cassette, was injected into the perivitelline space of murine fertilized oocytes before transplanting into the oviducts of the foster mothers. The GFP transgenic pups were then obtained. By PCR amplification, fluorescent microscopy and flow assisted cytometry sorting analysis, we found that the integration rate of the transgene was estimated at above 40%. Real-time PCR analysis indicated that the copy number of the integrated GFP cassette was around 40. Fluorescent in situ hybridization analysis demonstrated that the integration pattern was random but inheritable. The transgenic mice with multi-integration sites and various expression levels possessed a great value in practice as well as research. The approach reported herein provides an efficient way to generate and screen the transgenic mouse strains.
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| 关 键 词： | lentiviral vector transgene mouse integration GFP. |
Production of transgenic mice carrying green fluorescence proteingene by a lentiviral vector-mediated approach |
| |
| Authors: | ZHANG Jingzhi GUO Xinbing XIE Shuyang ZHU Yiwen HUANG Ying WANG Shu REN Zhaorui |
| |
| Institution: | Shanghai Institute of Medical Genetics, Shanghai JiaoTong University, Shanghai 200040, China |
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| Abstract: | A pseudo-lentivirus, which carries green fluorescence protein (GFP) expressing cassette, was injected into the perivitelline space of murine fertilized oocytes before transplanting into the oviducts of the foster mothers. The GFP transgenic pups were then obtained. By PCR amplification, fluorescent microscopy and flow assisted cytometry sorting analysis, we found that the integration rate of the transgene was estimated at above 40%. Real-time PCR analysis indicated that the copy number of the integrated GFP cassette was around 40. Fluorescent in situ hybridization analysis demonstrated that the integration pattern was random but inheritable. The transgenic mice with multi-integration sites and various expression levels possessed a great value in practice as well as research. The approach reported herein provides an efficient way to generate and screen the transgenic mouse strains. |
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| Keywords: | lentiviral vector transgene mouse integration GFP |
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