CAJ | 학술논문

目的：构建带Flag标签的MDM2真核表达载体，并检测MDM2与p53的相互作用。方法：从人乳腺文库中PCR扩增MDM2编码序列，将其插入pcDNA3.0-Flag载体，转染293T细胞后用Western印迹检测其在293T细胞中的表达，并通过免疫共沉淀实验检测MDM2与p53的相互作用。结果：双酶切和测序结果表明，Flag-MDM2真核表达载体构建成功，转染293T细胞后成功表达；免疫共沉淀实验证明Flag-MDM2与p53存在相互作用。结论：构建了带Flag标签的人MDM2真核表达载体，并检测了MDM2与p53之间的相互作用，为研究MDM2的功能奠定了基础。
목적：구건대Flag표첨적MDM2진핵표체재체，병검측MDM2여p53적상호작용。방법：종인유선문고중PCR확증MDM2편마서렬，장기삽입pcDNA3.0-Flag재체，전염293T세포후용Western인적검측기재293T세포중적표체，병통과면역공침정실험검측MDM2여p53적상호작용。결과：쌍매절화측서결과표명，Flag-MDM2진핵표체재체구건성공，전염293T세포후성공표체；면역공침정실험증명Flag-MDM2여p53존재상호작용。결론：구건료대Flag표첨적인MDM2진핵표체재체，병검측료MDM2여p53지간적상호작용，위연구MDM2적공능전정료기출。
Objective: To construct the eukaryotic expression vector of human MDM2 labeled with Flag tag and detect the interaction with p53. Methods: Human MDM2 gene was obtained from human mammary cDNA library by PCR and cloned into Flag vector. Human 293T cells were transfected with the recombinant plasmid of Flag-MDM2 and the expression was detected by Western blot. Interaction between MDM2 and p53 was detected by im-munoprecipitation. Results: MDM2 eukaryotic expression vector labeled with Flag tag was successfully constructed. The expression of Flag-MDM2 was identified by Western blot in human 293T cells. Flag-MDM2 interacted with p53. Conclusion: The eukaryotic expression vector of Flag-MDM2 was successfully constructed and interacted with p53, which had laid foundation for the further study of the function of MDM2.