心脏组织特异性表达Cre重组酶转基因小鼠的建立  

Establishment of cardiac-specific Cre recombinase transgenic mice

作　　者：王剑[1] 张莉[1] 毛春明[1] 程萱[1] 侯宁[1] 吕娅歆[1] 杨晓[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《军事医学科学院院刊》2005年第1期38-40,44,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家杰出青年基金(30128013 );国家自然科学基金面上项目(30070837)

摘　　要：　目的:建立在小鼠心肌细胞中特异性表达Cre重组酶的转基因小鼠。方法:构建了心肌细胞特异性表达Cre重组酶的转基因载体。该载体通过显微注射被导入小鼠受精卵中,通过胚胎移植,获得转基因首建者小鼠。利用PCR检测子代小鼠Cre重组酶整合情况。通过Northern杂交检测Cre重组酶表达的组织特异性。结果:构建了含有心肌细胞特异性α- 肌球蛋白重链(α- MHC)启动子、Cre重组酶基因和人生长激素基因polyA的转基因载体α- MHC -Cre -hGH。将转基因载体进行显微注射,共注射了 215枚小鼠受精卵,其中 202枚移植入 13只假孕母鼠的输卵管中发育,获得子代小鼠 42只。PCR检测发现有 1只小鼠在其基因组上整合有Cre重组酶基因。Northern杂交检测结果显示该转基因小鼠只在心脏组织中特异性地表达Cre重组酶基因。结论:成功获得了在小鼠心肌细胞中特异性表达Cre重组酶的转基因小鼠,为利用条件基因打靶技术研究基因在心脏发育与相关疾病中的功能提供了有利的工具。Objective: To obtain a mouse that specifically expresses Cre recombinase in cardiomyocyte. Methods:A construct that directed the expression of Cre recombinase to cardiomyocytes was made. The linearized constructs were introduced into the fertilized eggs by microinjection to obtain the transgenic mice. The integration of Cre recombinase gene was detected using PCR. The tissue-specific expression of the Cre recombinase was confirmed by Northern blot. Results:A cardiomyocyte-specific Cre transgenic construct containing the α-myosin heavy chain (α-MHC) promoter, Cre recombinase gene and polyA of human growth hormone gene was generated. The linearized constructs were introduced into 215 fertilized zygotes by microinjection. Two hundred and two injected eggs were implanted into the oviduct of 13 female mice. Forty two offspring were obtained. One mouse carrying the Cre recombinase gene was identified by PCR. The results of Northern blot examination of multiple tissues showed that the Cre recombinase was expressed only in the cardiac tissue. Conclusion: A cardiomyocyte-specific Cre recombinase transgenic mouse was generated successfully. It could serve as a useful tool for studying gene function in heart development and related diseases using conditional gene knockout approach.

关 键 词：心脏 CRE重组酶 小鼠 转基因 组织特异性 

分 类 号：Q813[生物学—生物工程]