AML1-ETO转基因小鼠的建立及初步表型分析

Establishment and phenotypic analysis in transgenic mice with AML1-ETO fusion gene

目的:建立AM L1-ETO融合基因转基因小鼠,在整体动物水平研究AM L1-ETO融合蛋白在白血病发病中所起的作用。方法:构建hCG/AM L1-ETO转基因质粒,通过显微注射将该质粒转入小鼠受精卵中,植入假孕母鼠输卵管,获得G0代转基因小鼠;用聚合酶链反应(PCR)方法检测融合基因的整合情况,逆转录-聚合酶链反应(RT-PCR)方法检测融合基因的表达情况和组织表达谱。结果:PCR法共检出10只G0代转基因阳性小鼠,其中8个系均产下F1代小鼠,由此建立了8个AM L1-ETO转基因小鼠系。RT-PCR法证实AM L1-ETO融合基因在22系中稳定表达,但血常规、肝脏、脾脏等组织未见异常变化;对该系小鼠的组织表达谱检测表明,融合基因在肝脏、脾脏、心脏和肌肉组织存在不同程度的表达,但在脑组织中不表达。结论:AM L1-ETO融合基因能在转基因小鼠的骨髓等组织中表达,但尚不足以引发白血病,推测其他致病基因在小鼠白血病的发生中也起一定作用。

Objective To investigate the leukemogenic potential of AML1-ETO fusion gene in vivo, and to provide an ideal animal model for anti-leukemia drug screening. Methods The molecular cloning technology was used to construct the AML1-ETO transgenic plasmid. The recombined DNA was microinjected into the zygotes, which was then implanted into the pseudopregnant mice. And then the G0 transgenic mice were obtained. The integration of the transgene was tested by PCR and the expression was examined by reverse transcription-polymerase chain reaction (RT-PCT). Results PCR testing revealed a total of 10 G0 transgenic mice, 8 of whom had their F1 offsprings and then 8 AML1-ETO transgenic mouse lines was established. RT-PCR showed the AML1-ETO fusion gene stably expressed only in one line, which had no abnormal changes in the blood routine, liver and spleen tissues. The AML1-ETO gene was found to express stably in liver, spleen, heart and muscle except brain. Conclusions A transgenic mouse model in which the AML1-ETO fusion gene can stably express has been established.