南方医科大学学报2016，Vol.36Issue(12)：1592-1598,7.DOI:10.3969/j.issn.1673-4254.2016.12.01
琼脂磁珠消减SELEX技术筛选HIV gp41抗原适配体
Subtractive SELEX using agar beads for screening DNA aptamers with specific affinity to HIV gp41 antigen
摘要
Abstract
Objective To obtain DNA aptamers with a highly specific affinity to HIV gp41 antigen using SELEX screening for detection of HIV.Methods The specific DNA aptamers of HIV gp41 antigen were screened from the double-stranded DNA derived from the single-stranded DNA (ssDNA) library with agarose beads as the supportive medium and HIV gp41 antigen as the target molecule using SELEX technique and real-time quantitative PCR.Results The secondary ssDNA library obtained after 6 rounds of screening was amplified by PCR to obtain dsDNA.The dsDNA was linked with pMDTM 18-T vector,cloned and sequenced to obtain 4 aptamers of HV gp41 antigen.The affinities of the 4 aptamers (Kd) all reached the nanomolar level.Among the 4 aptamers,the No.15 aptamer showed the strongest affinity.Specificity analysis of the aptamers revealed that all these 4 aptamers had specific affinity to HIV gp41 antigen with no affinity to other non-specific proteins.Conclusion We successfully obtained DNA aptamers with highly specific affinity to the HIV gp41 antigen from random single-stranded oligonucleotide library,and the obtained aptamers have the ability to antagonize HIV gp41 antigen.关键词
HIV gp41抗原/消减SELEX技术/DNA适配体识别Key words
HIV gp41 antigen/subtractive SELEX technique/DNA aptamers引用本文复制引用
栗坤,修尘林,高立明,石明,翟越..琼脂磁珠消减SELEX技术筛选HIV gp41抗原适配体[J].南方医科大学学报,2016,36(12):1592-1598,7.基金项目
秦皇岛市科学技术研究与发展计划(201501B034,201502A193)Supported by Qinhuangdao Science and Technology Research and Development Plan (201501B034,201502A193). （201501B034,201502A193）
