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目的:研究重组腺病毒载体Ad5/F35-HF2S在慢性粒细胞白血病K562细胞中的定位及与融合蛋白BcrAbl的相互作用。方法:将Ad5/F35-HF2S腺病毒载体感染K562细胞,然后RT-PCR和蛋白质印迹法鉴定其表达情况,采用免疫荧光和蛋白质印迹法鉴定融合蛋白HF2S在K562细胞中的定位,免疫荧光法检测HF2S与Bcr-Abl在细胞质中的共定位情况,免疫共沉淀技术验证二者之间是否存在相互作用。结果:重组腺病毒Ad5/F35-HF2S能高效感染白血病K562细胞,并进行了正确表达。重组蛋白HF2S与Bcr-Abl共定位于细胞质,并能相互结合。结论:在K562细胞质中成功表达能与Bcr-Abl相互作用的HF2S蛋白,为后续靶向转运Bcr-Abl入核并治疗慢性粒细胞白血病奠定了基础。
AIM: To investigate the localization of recombinant adenovirus vector Ad5 / F35-HF2S in K562 cells and its interaction with BcrAb1 fusion protein. Methods: Ad5 / F35-HF2S adenovirus vector was used to infect K562 cells. The expression of fusion protein HF2S in K562 cells was identified by RT-PCR and Western blotting. The immunofluorescence assay The co-localization of HF2S and Bcr-Abl in the cytoplasm, co-immunoprecipitation technique to verify the existence of the interaction between the two. Results: Recombinant adenovirus Ad5 / F35-HF2S efficiently infected leukemia K562 cells and correctly expressed. Recombinant protein HF2S and Bcr-Abl co-localized in the cytoplasm, and can combine with each other. Conclusion: The successful expression of HF2S protein interacting with Bcr-Abl in the cytoplasm of K562 lays the foundation for the subsequent targeting of Bcr-Abl into the nucleus and the treatment of chronic myeloid leukemia.