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目的建立Bcl-2高表达的转基因小鼠品系,为在活体动物上研究bcl-2蛋白的功能提供动物模型。方法构建含人Bcl-2基因的质粒;通过显微注射将该质粒转入C57BL/6J×CBA的杂交小鼠的受精卵,并植入假孕母鼠的子宫,PCR检测后携带人Bcl-2基因的小鼠即为“Founder”鼠;将“Founder”鼠与C57BL/6J×CBA杂交小鼠不断回交,经PCR和Western bilotting筛选后即可获得阳性小鼠。结果共获得5只“Founder”鼠,其中#6、#14成功建系,连续回交繁殖7代后,PCR和Western筛选后获得bcl-2高表达阳性小鼠42只。结论成功将人bcl-2基因转入小鼠基因组内,并稳定传代,建立Bcl-2高表达小鼠,为进一步研究Bcl-2蛋白提供了很好的动物模型。
OBJECTIVE: To establish a transgenic Bcl-2-overexpressing mouse strain that provides an animal model for studying the function of bcl-2 protein in living animals. Methods Plasmid containing human Bcl-2 gene was constructed. The plasmid was transfected into the fertilized egg of C57BL / 6J × CBA hybrid mice by microinjection and implanted into the uterus of pseudopregnant female rats. After PCR, the plasmid carrying human Bcl- 2 mice were “Founder ” mice. The “Founder ” mice and C57BL / 6J × CBA hybrid mice were continuously backcrossed, and positive mice were screened by PCR and Western bilotting. Results A total of 5 “Founder” mice were obtained. Among them, # 6 and # 14 were successfully established. After 7 generations of continuous backcross breeding, 42 mice with high expression of bcl-2 were obtained after PCR and Western blotting. Conclusion The successful cloning of human bcl-2 gene into the mouse genome and the stable passage of Bcl-2 gene in mice resulted in the establishment of a Bcl-2-overexpressing mouse model, which provided a good animal model for the further study of Bcl-2 protein.