北华大学学报（自然科学版）2025，Vol.26Issue(1)：49-54,6.DOI:10.11713/j.issn.1009-4822.2025.01.008
盐酸胍法构建骨肉瘤细胞适配体ssDNA文库可行性
Feasibility of Guanidine Hydrochloride Method for ssDNA Library Construction of Osteosarcoma Aptamer
摘要
Abstract
Objective Exploring feasibility of Streptavidin-Biotin(STV-Biotin)system coupled with Guanidine hydrochloride(G-HCl)method for construction of an osteosarcoma cell aptamer ssDNA library.Methods The candidate oligonucleotide library ssDNA was selectively bound to target cell(osteosarcoma cell U2-OS),and the oligonucleotides were amplified into double-stranded DNA(dsDNA)by PCR,dsDNA were labeled with biotin and combined with Streptavidin Sepharose(S.S)affinity chromatography.The target single-stranded DNA(ssDNA)library was prepared by G-HCl method and purified.The classical NaOH method was used for control.The single strand libraries prepared by the two methods were compared with osteosarcoma cells for compatibility and recovery.Results Screening library ssDNA obtained by G-HCl method have higher affinity to U2-OS and higher recovery of ssDNA than classical NaOH method.Conclusion STV-Biotin system coupled with G-HCl is a successful attempt for the preparation of osteosarcoma cell aptamer,which provides a new idea and method for the const-ruction of tumor aptamer ssDNA library.关键词
适配体/骨肉瘤细胞/盐酸胍/PCR/单链寡核苷酸Key words
aptamer/osteosarcoma cells/Guanidine hydrochloride/PCR/ssDNA分类
医药卫生引用本文复制引用
魏唐文,魏兵,黄丹菊,梁昭珺,张宇飞,王秀娟,傅桂莲,梁骥..盐酸胍法构建骨肉瘤细胞适配体ssDNA文库可行性[J].北华大学学报（自然科学版）,2025,26(1):49-54,6.基金项目
广西自然科学基金项目(2018GXSFAA138035) （2018GXSFAA138035）
大学生创新创业训练计划项目(国家级,202310601054 （）
校级,202210601237) （）
广西高校中青年教师科研基础能力提升项目(2022KY0511) （2022KY0511）
广西肿瘤免疫与微环境调控重点实验室开放课题(2202KF004). （2202KF004）
