Isolation and Characterization of a ssDNA Aptamer against Major Soluble Antigen of Renibacterium salmoninarum
Abstract
:1. Introduction
2. Results and Discussion
2.1. MSA Protein and Constructs Generation
2.2. Isolation of DNA Aptamer
2.3. Analysis of Aptamer Structures
2.4. Binding Affinity and Specificity Test
3. Materials and Methods
3.1. Reagents and Instrumentation
3.2. Cloning, Expression and Purification of MSA Protein and Sub Constructs
3.3. Random Oligonucleotide Library and Primers
3.4. In Vitro Selection via GO-SELEX
3.5. Cloning, Sequencing, and Structure Analysis of Selected Aptamers
3.6. Binding Assay Procedure
3.7. Specificity Test
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Name | DNA Aptamer Sequence | Frequency |
---|---|---|
Aptamer-1 | 5′-ATGATACGGCGACCACCTAACGCACACAATATATTAAGGATCAAATTAATACTCGGTATATAGCGTCGTGTAGGGAAAGA-3′ | 1 |
Aptamer-2 | 5′-ATGATACGGCGACCACCTAAACATATGGAGTATTAGGGTTTACATAATTATGTATTTCTCTAGCGTCGTGTAGGGAAAGA-3′ | 1 |
Aptamer-3 | 5′-ATGATACGGCGACCACCTAAATTACTACATTAATTAGTTGTATAATTGTAGTATGATGATTAGCGTCGTGTAGGGAAAGA-3′ | 2 |
Aptamer-4 | 5′-ATGATACGGCGACCACCTAACCATTTTCACTGTTTGTTTAATATATATAGTTTTATGGTATAGCGTCGTGTAGGGAAAGA-3′ | 6 |
Aptamer-5 | 5′-ATGATACGGCGACCACCTAAAGTTATTTTTGTTATTATAACGGGTTACCTTTATAGACTATTAGCGTCGTGTAGGGAAAGA-3′ | 1 |
Aptamer-6 | 5′-ATGATACGGCGACCACCTAATTTATTAACTATTAACTGTTTAGGTGAGACTTTTCTGTTGTAGCGTCGTGTAGGGAAAGA-3′ | 1 |
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Layman, B.; Mandella, B.; Carter, J.; Breen, H.; Rinehart, J.; Cavinato, A. Isolation and Characterization of a ssDNA Aptamer against Major Soluble Antigen of Renibacterium salmoninarum. Molecules 2022, 27, 1853. https://doi.org/10.3390/molecules27061853
Layman B, Mandella B, Carter J, Breen H, Rinehart J, Cavinato A. Isolation and Characterization of a ssDNA Aptamer against Major Soluble Antigen of Renibacterium salmoninarum. Molecules. 2022; 27(6):1853. https://doi.org/10.3390/molecules27061853
Chicago/Turabian StyleLayman, Brady, Brian Mandella, Jessica Carter, Haley Breen, John Rinehart, and Anna Cavinato. 2022. "Isolation and Characterization of a ssDNA Aptamer against Major Soluble Antigen of Renibacterium salmoninarum" Molecules 27, no. 6: 1853. https://doi.org/10.3390/molecules27061853
APA StyleLayman, B., Mandella, B., Carter, J., Breen, H., Rinehart, J., & Cavinato, A. (2022). Isolation and Characterization of a ssDNA Aptamer against Major Soluble Antigen of Renibacterium salmoninarum. Molecules, 27(6), 1853. https://doi.org/10.3390/molecules27061853