| Issue |
BIO Web Conf.
Volume 8, 2017
2016 International Conference on Medicine Sciences and Bioengineering (ICMSB2016)
|
|
|---|---|---|
| Article Number | 03009 | |
| Number of page(s) | 10 | |
| Section | Session III: Biomedical Engineering | |
| DOI | https://doi.org/10.1051/bioconf/20170803009 | |
| Published online | 11 January 2017 | |
Screening ssDNA aptamers against HIV P24 antigen using agarose beads as carriers
1 College of Life Sciences, Northwest Normal University, Lanzhou, 730070, China
2 Laboratory of Molecular Biology, Gansu Academy of Medical Sciences, Lanzhou, 730050, China
a Corresponding author
In this study, carboxylated agarose beads were used as screening carriers combined with the subtractive systematic evolution of ligands by exponential enrichment (SELEX) technology to obtain an ssDNA secondary library, which could specifically bind to HIV P24 antigen after nine screening rounds from a random ssDNA library. An electrophoretic mobility shift assay (EMSA) and quantitative real-time PCR assay (RT-qPCR) was performed to monitor in real-time the binding specificity of the ssDNA secondary library to the HIV P24 antigen. The obtained ssDNA secondary library was converted to the corresponding dsDNA library, ligated to a PMD18-T vector, and transformed into E. coli DH5α and sequenced. The results demonstrated that after nine rounds of screening, two aptamer sequences specifically bound to the HIV P24 antigen were obtained.
© The Authors, published by EDP Sciences, 2017
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