摘要
目的:检测Cre重组酶在心肌细胞特异性Cre重组酶转基因小鼠(α-MHC-Cre)中的组织分布及其在体内介导基因重组的作用。方法:将α-MHC-Cre转基因小鼠与ROSA26报告小鼠交配,利用LacZ染色对双转基因阳性子代小鼠进行检测。然后,将α-MHC-Cre小鼠与Sm ad4条件基因打靶小鼠交配,利用PCR和Southern杂交对Cre重组酶介导重组的组织特异性进行检测。结果:LacZ染色表明,Cre重组酶只在心肌细胞中特异性表达并介导ROSA位点LoxP序列间的重组。PCR和Southern结果显示Cre重组酶只在心肌细胞中特异地剔除了Sm ad4基因,进一步验证了Cre重组酶在心肌细胞中发挥介导LoxP位点重组的作用。结论:α-MHC-Cre转基因小鼠具有良好的组织特异性,只在心肌细胞中表达Cre重组酶,并能在体内成功地介导心肌细胞基因组上LoxP位点间的重组,是一种理想的研制心肌细胞特异性基因剔除小鼠的工具小鼠。
Objective： To determine the tissue distribution and the activity of Cre recombinase in the cardiomyocyte-specific Cre transgenic mouse line expressing the Cre recombinase under the control of mouse α-MHC gene promoter. Methods： Firstly, the α-MHC-Cre transgenic mice were mated with ROSA26 reporter strain. The activity of Cre recombinase was examined by LacZ staining in the double transgenic mice. Then the α-MHC-Cre transgenic mice were mated with a mouse strain that carried Smad4 conditional alleles （Smad4^Co/Co）. PCR and Southern blot were used to examine the Cre mediated recombination in different tissues of the offsprings. Results： LacZ staining showed that Cre recombinase was expressed and functioned only in the cardiac tissue, especially in cardiomyocytes. The result was further verified by PCR and Southern blot,which revealed that the Smad4 gene was deleted only in the cardiac tissue. Conclusion： All these data indicated that the Cre recombinase was expressed exclusively in the cardiac tissues of the α-MHC-Cre transgenic mice. The α-MHC-Cre transgenic mice could serve as a useful tool for generating cardiomyocyte-specific gene-knockout mice.
出处
《军事医学科学院院刊》
CSCD
北大核心
2005年第5期447-449,456,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)
