HIF-1α条件性基因敲除嵌合体小鼠的获得  

Generation of chimeras for HIF-1α conditional knockout mice

作　　者：赵玲[1] 杜晓兰[1] 苏楠[1] 宋瑞华[1] 何启芬[1] 李福兵[1] 戚华兵[1] 陈林[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所全军创伤中心创伤,烧伤与复合伤国家重点实验室,重庆400042

出　　处：《第三军医大学学报》2007年第14期1361-1363,共3页Journal of Third Military Medical University

基　　金：国家重点基础研究发展规划项目("973"项目)(G1999054203;2005CB522604);国家自然科学基金面上项目(30370318)~~

摘　　要：目的应用Cre/loxP系统建立HIF-1α条件性基因敲除小鼠模型。方法电击法将构建好的HIF-1α条件性基因敲除载体转染入小鼠ES细胞,用G418与Ganciclovir对转染的ES细胞进行正负筛选,经Southern杂交鉴定得到阳性ES细胞克隆后,将已发生正确重组的ES细胞显微注射入小鼠囊胚腔,对子代鼠进行基因型鉴定。结果经G418与Gan-ciclovir筛选培养得到490个阳性克隆,用5′端探针进行Southern杂交鉴定获得1株发生同源重组的ES细胞克隆,经显微注射,得到5只由基因敲除ES细胞和供体囊胚细胞共同发育而来的嵌合体小鼠。结论成功得到基于Cre/loxP系统的HIF-1α条件性基因敲除嵌合体小鼠模型。Objective To generate chimeras for hypoxia inducible factor-1 α （HIF-1 α） conditional knockout mice using Cre/loxP system. Methods The mouse ES cells were electroporated with conditional knockout vector of HIF-1 α, selected by G418-positive and Ganciclovir-negative culture. Target ES ceils, following positive and negative selection, was identified by Southern blotting. Chimera mice were born by pseudopregenant mice which were transplanted with blastocysts microinjected with target ES cells. Results A total of 490 ES clones were collected after screening by both G418-positive and Ganciclovir-negative selection, and one clone with correct homologous recombinant event was obtained by Southern blotting screening. Five chimeras were produced following blastocysts microinjection and transplantation. Conclusion Chimera mice with HIF-1α conditional knockout allele were obtained, which will help to finally obtain HIF-1 α conditional knockout mice, subsequently provide the possibility to study HIF-1 α function in vivo.

关 键 词：低氧诱导因子-1Α 条件性基因敲除 同源重组 胚胎干细胞 CRE/LOXP系统 

分 类 号：R-332[医药卫生] R394-33