基础医学与临床 ›› 2012, Vol. 32 ›› Issue (9): 987-991.

• 研究论文 •    下一篇

HIV-p24核酸适配体的筛选

张宁1,毛爱红2,马瑾2,廖世奇2,张丽琼1,王晓清2,马君1   

  1. 1. 兰州理工大学
    2. 甘肃省医学科学研究院
  • 收稿日期:2011-08-16 修回日期:2011-12-02 出版日期:2012-09-05 发布日期:2012-08-28
  • 通讯作者: 廖世奇 E-mail:lshiqi@126.com
  • 基金资助:
    骨髓间充质干细胞瘤化中STAT3信号通路过度激活的作用机制与规避研究;骨髓间充质干细胞瘤化中STAT3信号通路过度激活的作用机制与规避研究;骨髓间充质干细胞瘤化中STAT3信号通路过度激活的作用机制与规避研究;国家中小企业创新基金;甘肃省重大研发项目;甘肃省重大研发项目

Screening of aptamers to HIV-p24

  • Received:2011-08-16 Revised:2011-12-02 Online:2012-09-05 Published:2012-08-28

摘要: 摘 要:目的 利用SELEX技术筛选HIV-p24的核酸适配体,为艾滋病的诊断和治疗奠定基础。方法 以重组p24为筛选靶,用SELEX技术从随机寡核苷酸库中筛选与HIV-p24结合的寡核苷酸,利用凝胶阻滞实验鉴定第12轮筛选到的寡核苷酸与HIV-p24的结合,再用Dot-blot法筛选出与HIV-p24结合的核酸适配体,并检测核酸适配体识别HIV-p24的特异性。结果 Dot-blot筛选到5条与HIV-p24有较强结合能力的核酸适配体,且均为不同的序列。特异性检测显示,18和26号配体只与HIV-p24特异性结合,与人血清白蛋白、牛血清白蛋白和脱脂奶粉均无明显结合。结论 成功筛选到2条特异结合HIV-p24的核酸适配体,为其应用于艾滋病诊断和治疗提供了实验基础。

关键词: 关键词:SELEX技术, HIV-p24, 核酸适配体

Abstract: Abstract: Objective Aptamers to HIV-p24 were screened through SELEX technique for the diagnosis and therapy of AIDS. Methods With recombinant p24 for the screening target, oligonucleotides binding HIV-p24 were screened from a random oligonucleotide library through SELEX technique. The binding capacity between oligonucleotides obtained from the 12th round of screening and HIV-p24 was identified via Electrophoretic Mobility Shift Assay (EMSA).The aptamers strongly binding to HIV-p24 were screened and the specificity of aptamers recognizing HIV-p24 was detected by Dot-blot method. Results 5 aptamers with strong binding capacity to HIV-p24 were obtained and are different sequences. The binding speci?city indicated that No.18 and No.26 apatmers only bond with HIV-p24, not with human serum albumin, bovine serum albumin and skimmed milk powder. Conclusion 2 aptamers specially binding to HIV-p24 were obtained, this study provides an experimental basis for the diagnosis and treatment of AIDS by utilizing aptamer of HIV-p24.

Key words: Key words: SELEX technique, HIV-p24, aptamer

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