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目的 :研究特异识别已分化PC12细胞的ssDNA适体的二级结构及截短前后的适体与已分化PC12细胞的结合能力。方法 :利用RNAstructure 3.5软件对截短前后的适体的空间结构进行模建 ,体外合成这些截短适体 ,利用流式细胞术和荧光显微镜研究适体与已分化PC12细胞的结合能力。结果 :单茎环结构是适体与已分化PC12细胞结合的二级结构 ;截短后的适体与已分化PC12细胞的结合能力明显大于与未分化PC12细胞结合能力 ;AP17 38与已分化PC12细胞结合的荧光强度明显大于与未分化PC12细胞结合的荧光强度 ;截短前后的适体均能够特异识别混合物中已分化PC12细胞。结论 :截短后的适体能够保留适体的空间结构 ,而且与已分化PC12细胞的亲合力不会降低。
OBJECTIVE: To study the secondary structure of ssDNA aptamers that specifically recognize differentiated PC12 cells and the binding ability of aptamers to differentiated PC12 cells before and after truncation. METHODS: RNAstructure 3.5 software was used to construct the aptamer spatial structure before and after truncation. These truncated aptamers were synthesized in vitro. The binding ability of aptamers to differentiated PC12 cells was studied by flow cytometry and fluorescence microscopy. Results: The single stem-loop structure was the secondary structure of the aptamer and the differentiated PC12 cells. The binding capacity of the truncated aptamer to the differentiated PC12 cells was significantly higher than that of the undifferentiated PC12 cells. The intensity of cell-bound fluorescence was significantly greater than that of undifferentiated PC12 cells; both aptamer before and after truncation were able to specifically recognize differentiated PC12 cells in the mixture. CONCLUSION: The truncated aptamer retains the aptamer’s spatial structure, and its affinity with differentiated PC12 cells does not decrease.