Subtractive SELEX using agar beads for screening DNA aptamers with specific affinity to HIV gp41 antigen
LI Kun
XIU Chenlin
GAO Liming
SHI Ming
ZHAI Yue
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.
Keywords：HIV gp41 antigensubtractive SELEX techniqueDNA aptamers
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 1592-1598 )