| 小鼠胚胎成纤维细胞的转染与阳性细胞的初步筛选 |
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| 引用本文： | 姜秋,倪雪岩,聂代邦,孙宏晨. 小鼠胚胎成纤维细胞的转染与阳性细胞的初步筛选[J]. 吉林大学学报(医学版), 2008, 34(6): 990-993. DOI: 吉林省科技厅科技发展计划项目资助课题（20 |
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| 作者姓名： | 姜秋 倪雪岩 聂代邦 孙宏晨 |
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| 作者单位： | 吉林大学口腔医院儿童口腔科,吉林,长春,130041;首都医科大学附属北京安贞医院口腔医疗中心,北京,100029;吉林大学畜牧兽医学院生物技术系,吉林,长春,130062 |
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| 基金项目： | 吉林省科技厅科技发展计划项目资助课题 |
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| 摘 要： | 目的：建立表达白血病抑制因子（LIF）的胚胎成纤维细胞的方法体系，为建立转基因动物模型提供理论依据。方法：以13.5 d ICR小鼠胚胎为材料，采用胰酶消化方法，培养小鼠原代胚胎成纤维细胞。利用脂质体介导方法，将线性化的pcDNA3.1-LIF真核表达载体转染至小鼠胚胎成纤维细胞中。经G418筛选阳性细胞，提取阳性细胞的基因组DNA并测序。结果：通过对小鼠原代胚胎成纤维细胞的分离、培养，获得原代胚胎成纤维细胞。经过转染，建立了表达LIF的胚胎成纤维细胞,并在倒置显微镜下观察到筛选得到的阳性细胞。对阳性细胞克隆质粒测序表明同源性达99%。结论：pcDNA3.1-LIF真核表达载体成功转染到小鼠胚胎成纤维细胞中。
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| 关 键 词： | 白血病抑制因子 成纤维细胞 转染 小鼠 |
| 收稿时间： | 2007-11-21 |
Transfer of mouse fetal fibroblasts and primary selection of positive cells |
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| JIANG Qiu,NI Xue-yan,NIE Dai-bang,SUN Hong-chen. Transfer of mouse fetal fibroblasts and primary selection of positive cells[J]. Journal of Jilin University: Med Ed, 2008, 34(6): 990-993. DOI: 吉林省科技厅科技发展计划项目资助课题（20 |
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| Authors: | JIANG Qiu NI Xue-yan NIE Dai-bang SUN Hong-chen |
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| Affiliation: | （1.Department of Pedodontics，Stomatology Hospital，Jilin University，Changchun 130041，China；2. Department of Stomatology, Beijing Anzhen Hospital,Affiliated to Capital University of Medical Sciences,Beijing 100029,China;3．Department of Animal Biotechnology，School of Animal Science and Veterinary Medicine，Jilin University，Changchun 130062，China） |
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| Abstract: | Objective To establish the method to express leukemia inhibitory factor(LIF) in fetal fibroblasts in order to provide theoretical foundation for establishment of transgenic animal model.Methods Using the fetal of 13.5 d ICR mouse,the primary fetal fibroblasts were cultivated by trypsin enzyme digestion.The lineared eukaryotic expression vector pcDNA3.1-LIF was transferred to the fetal fibroblasts of mouse with liposome induction.The positive cells were selected by G418,genome DNA of the positive cells was e... |
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| Keywords: | leukemia inhibitory factor fetal fibroblasts transfer mice |
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