心肌α-MHC启动子驱动的人KIAA0174基因转基因小鼠的建立

Transgenic mice expressing human KIAA0174 gene in heart tissue under control of α-cardiac myosin heavy chain promoter

目的:建立心肌高表达人KIAA0174基因转基因小鼠,研究该基因在心肌表达水平的变化对动物的影响,探索该基因的功能。方法:RT-PCR法克隆人KIAA0174基因,把该基因插入心肌α肌球蛋白重链启动子下游,构建转基因表达载体,通过显微注射法建立心肌高表达人KIAA0174基因转基因C57BL/6J小鼠。并利用特异引物PCR法鉴定转基因小鼠的基因型,Western blot检测基因表达水平。测定小鼠的心率、收缩压,以及扩张压的平均值。结果:建立了3个心肌高表达人KIAA0174基因转基因小鼠系。血压检测结果显示转基因小鼠的收缩压和扩张压均显著高于对照组。结论:成功建立了心肌α肌球蛋白重链启动子驱动的心肌高表达人KIAA0174基因转基因小鼠,为进一步探索KIAA0174基因的功能奠定了基础。

Objective：To generate the heart specific KIAA0174 transgenic mice and investigate its functions.Methods：KIAA0174 gene was ligated down stream of alpha-cardiac myosin heavy chain promoter（alpha-MHC） to construct the heart specific vector.The transgenic mice were produced by microinjection method and the genotyping was detected by PCR.The expression levels of the transgenic gene were detected with Western blot.Blood pressure and heart rate in conscious mice were measured by the tail-cuff system.Results：Three lines of heart specific transgenic C57BL/6J mice of human KIAA0174 gene were established.Our results suggested a significant difference between transgenic and non-transgenic mice in systolic blood pressure（SBP） and diastolic blood pressure（DBP）.Conclusion：The transgenic mice that express the human KIAA0174 gene in heart tissue are established successfully.The establishment of the transgenic mice is essential for investigating the function of this gene.