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造血干细胞特异性表达Cre重组酶转基因小鼠的建立

DOI: 10.1360/052011-545, PP. 933-937

Keywords: Cre-Loxp重组系统,EDAG,转基因小鼠,组织特异性表达

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Abstract:

EDAG是在胚胎发育阶段造血干细胞特异性表达的基因.为了在早期造血组织细胞中实现相关基因的条件敲除,构建了含有早期造血组织特异性表达的EDAG启动子和Cre重组酶基因的转基因EDAG-Cre表达载体质粒.通过显微注射的方法将线性化的5.6kb的EDAG-Cre转基因片段导入小鼠受精卵细胞核,获得的新生小鼠经过PCR鉴定,常规方法培育传代.结果发现,共获得了6只阳性转基因首建鼠,其中4只已经建系并稳定传代.RT-PCR分析表明Cre重组酶基因在阳性转基因小鼠的骨髓、脾脏、胸腺、外周血以及胎肝等组织中均有表达,重组酶活性也在脾和骨髓中获得确认.EDAG-Cre重组酶转基因小鼠的建立,为研究早期造血组织以及造血干细胞特异性基因条件敲除小鼠模型的建立奠定了基础.

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