| 小鼠ADAM10条件性基因打靶载体的构建与鉴定 |
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| 引用本文： | 林炤华,杨丽华,郑杰辉,周常文. 小鼠ADAM10条件性基因打靶载体的构建与鉴定[J]. 中国优生与遗传杂志, 2009, 0(12): 17-19 |
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| 作者姓名： | 林炤华 杨丽华 郑杰辉 周常文 |
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| 作者单位： | 福建医科大学基础医学院细胞生物学与遗传学系细胞与发育工程研究中心,福州350004 |
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| 基金项目： | 福建医科大学科研发展基金（XZ04005） |
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| 摘 要： | 目的构建小鼠ADAM10条件性基因打靶载体,为建立ADAM10条件性敲除小鼠模型奠定基础。方法以正常小鼠（129/Svj）基因组DNA为模板,采用长片段PCR方法,扩增小鼠ADAM10基因第2外显子及其侧翼序列。通过引入LoxP位点和TK基因等步骤,获得ADAM10条件性基因打靶载体。结果经限制性内切酶酶切鉴定和测序证实,构建的小鼠ADAM10基因条件性打靶载体符合设计要求。结沦成功构建了小鼠ADAM10条件性基因打靶载体,为建立ADAM10条件性基因敲除小鼠打下了基础。
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| 关 键 词： | ADAM10 条件性基因打靶 同源重组 |
Construction and characterization of the targeting vector of conditional knockout of mouse ADAM10 gene |
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| Affiliation: | LIN Zhao - hua, YANG Li - hua, ZHENG Jie - hui, ZHOU Chang - wen. ( Department of Cell Biology and Genetics, Center of Cellular and Developmental Engineering, Fujian Medical University, Fuzhou, 350004) |
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| Abstract: | Objective： To construct the targeting vector of conditional knockout of ADAM10 gene for the generation of ADAM10 - deficient mouse. Methods： 6286bp and 6007bp genomic DNA fragments containing exon 2 of ADAM10 gene from 129/Svj mouse strain were amplified by PCR and inserted into T vector （pMD - 18T）, respectively, which were used as the homologous arms. Two LoxP sites were inserted into intron 1 and 2 for conditional knockout of exon 2 of ADAM10 gene. Results ： The correct structure of the targeting vector was confirmed by restriction endonuclease digestion and sequencing analysis. Conclusion： A targeting vector for conditional knockout of ADAM10 gene has been successfully constructed, which lay a foundation for generating the conditional ADAM10 - deleted mouse. |
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| Keywords: | ADAM10 Conditional gene targeting Homologous recombination |
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