| 敲除PKD1基因的小鼠胚胎干细胞的建立 |
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| 引用本文： | 郁胜强,梅长林,胡以平,王新民. 敲除PKD1基因的小鼠胚胎干细胞的建立[J]. 第二军医大学学报, 2003, 24(1): 5-10 |
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| 作者姓名： | 郁胜强 梅长林 胡以平 王新民 |
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| 作者单位： | 第二军医大学附属长征医院;第二军医大学附属长征医院 |
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| 基金项目： | This work is supported by the10 th Five YearPlan Program for Major Sci-tech Foundation (No.2 0 0 2 AA2 Z3 13 0 ),National Natural Science Foundation ofChina(No.3 0 170 90 1,No.3 0 2 715 2 3 ),The Hundred LeadingScientists Program of the Public |
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| 摘 要： | 目的：通过PKD1基因打靶载体的胚胎干细胞转染,药物筛选以及分子鉴定,获得发生同源重组的胚胎干细胞克隆,为产生PKD1基因缺陷小鼠品系准备条件。方法：体外培养小鼠胚胎干细胞,用电穿孔的方法进行PKD1基因打靶载体的转移,并用G418进行筛选培养,得到阳性克隆后,进一步扩大培养,抽提胚胎干细胞的基因组DNA,采用DNA印迹法进行分子鉴定。结果：经G418筛培养得到192个阳性在隆,分子鉴定获得2个发生同源重组的胚胎干细胞克隆。结论：鉴定得到的2个同源重组胚胎干细胞克隆,为进一步建立PKD1基因缺陷小鼠奠定了基础。
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| 关 键 词： | 胚胎干细胞 多囊肾病 基因敲除 |
Establishing embryonic stem cells model in PKD1 knock-out mouse |
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| Abstract. Establishing embryonic stem cells model in PKD1 knock-out mouse[J]. Former Academic Journal of Second Military Medical University, 2003, 24(1): 5-10 |
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| Authors: | Abstract |
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| Abstract: | Objective:To obtain the homologous recombinant embryonic stem cells of the mouse PKD1 gene targeting vector.Methods:Mouse embryonic stem cells were cultured and observed in vitro,after transfected with targeting vector DNA (linearized with XbaⅠ) by electroporation,some ES cell lines survived G418 selection.G418 resistant colonies were expanded and identified by Southern blotting.Results:Totally 192 ES cell lines survived G418 selection and homologous recombinant colonies were obtained.Conclusion:The results of this experiment are the foundation of the PKD1 transgenic mouse model. |
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| Keywords: | embryonic stem cells polycystic kidney disease gene knock out |
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