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目的 建立 HRX- EEN融合基因转基因小鼠为新药的筛选提供理想的动物模型。 方法 通过拼接 ,用 3个不同来源的 DNA构建成 HRX- EEN融合基因 ,并将其装入 h CG转基因载体中 ;通过显微注射将上述 DNA导入受精卵雄核中 ,植入养育母鼠输卵管 ,发育获得 G0 代小鼠 ;PCR检测融合基因的整合 ,逆转录 -聚合酶链反应 (reverse transcription- polymerase chain reaction,RT- PCR)检测融合基因的表达。结果 测序证实拼接的 HRX- EEN融合基因正确 ;PCR共检测出 7个 HRX- EEN转基因阳性的 G0 代小鼠 ,将这些小鼠与野生型的 C5 7系小鼠交配 ,结果除 35号死亡外 ,其余 G0 代转基因阳性小鼠均产下 F1代 ,建成 6个单系的 HRX- EEN转基因小鼠。用 RT- PCR方法检测其中 5个系小鼠 HRX- EEN融合基因的表达状况 ,证实该融合基因在 3个单系中稳定表达。表达融合基因的转基因小鼠迄今未发现明显异常。 结论 构建完成的 HRX- EEN转基因小鼠模型能稳定表达。
Objective To establish a transgenic mouse model of HRX-EEN fusion gene to provide an ideal animal model for the screening of new drugs. METHODS: The spliced HRX-EEN fusion gene was constructed from three different DNA sources and inserted into the h CG transgene vector. The DNA was introduced into the nucleus of the fertilized egg by microinjection, G0 mice were obtained. The integration of fusion gene was detected by PCR and the expression of fusion gene was detected by reverse transcription-polymerase chain reaction (RT-PCR). The results of sequencing confirmed that the spliced HRX-EEN fusion gene was correct; PCR detected seven HRX-EEN transgenic positive G0 mice and crossed these with wild-type C5 7 mice. , The remaining G0 generation of transgenic positive mice were produced F1 generation, built 6 single line of HRX-EEN transgenic mice. The expression of HRX-EEN fusion gene in 5 lines of mice was detected by RT-PCR, and it was confirmed that the fusion gene was stably expressed in 3 lines. So far, no significant abnormality has been found in the transgenic mice expressing the fusion gene. Conclusion The constructed HRX-EEN transgenic mouse model can stably express.