摘要
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.
EDAG is a specifically expressed gene of hematopoietic stem cell during embryo development.To obtain hematopoietic stem cell specific Cre recombinase transgenic mice,and construct conditional knocking-out target gene during early hematopoiesis,a targeting vector（EDAG-Cre） containing EDAG promoter and Cre recombinase gene was generated and linearized.By microinjecting 5.6 kb linearized EDAG-Cre into pronuclear fertilized oocyte,the pups were characterized by genomic PCR,and the conventional method was used for proliferation.The results show that six Cre positive transgenic mice were identified,and four of them were stably transferred.RT-PCR analysis shows that the Cre recombinase gene was expressed in bone marrow,spleen,thymus,peripheral blood,fetal liver and other tissues of the positive transgenic mice,and Cre recombinase activity in bone marrow and spleen were also confirmed.It demonstrates that EDAG-Cre transgenic mice were established and could be used for generating hematopoietic stem cell specific conditional knockout mice.
出处
《中国科学：生命科学》
CSCD
北大核心
2011年第10期933-937,共5页
Scientia Sinica(Vitae)
基金
国家自然科学基金(批准号:30871416)
北京市自然科学基金(批准号:5092021)
教育部留学回国人员科研启动基金(批准号:2010-2012)资助项目
