表达SARS冠状病毒受体人ACE2转基因小鼠的建立

Establishment of transgenic mice which express SARS coronavirus functional receptor human ACE2

目的:建立表达SARS冠状病毒的功能性受体人ACE2的转基因小鼠。方法:克隆人2型血管紧张素转换酶(hACE2)的cDNA,连入PCAGGS构建转基因表达载体,以显微注射的方法将5.6kb的转基因片段引入小鼠的受精卵。采用PCR、Southern印迹、RT-PCR、Western印迹、免疫荧光染色的方法对转基因小鼠进行鉴定。结果与结论:获得了在小鼠肺组织中表达有SARS病毒功能性受体hACE2的两个转基因小鼠系,证明了SARS病毒的S蛋白能很好地和转基因小鼠肺组织中表达的hACE2受体结合,这将为SARS的研究提供一种良好的感染动物模型。

Objective： To establish transgenic mice that express severe acute respiratory syndrome associated coronavirus （SARS-Cov） functional receptor human angiotensin converting enzyme type2 （ hACE2 ）. Methods： hACE2 eDNA was cloned and the expression vector PCAGGS-hACE2 was constructed. The 5.6 kb transgenic DNA fragments were introduced into mice fertilized eggs by microinjection, RCR, Southern blot, RT-PCR, Western blot and immunofluorescence staining were used to examine the integration and the expression of the exogenous hACE2 gene. Results and Conclusion： Two transgenic mouse lines that express the human ACE2 were obtained, and the hACE2 expressed in the mice lung can bind the S protein of SARS-Cov. It will provide an animal model for the research on SARS-Cov infection.