ArticleNucleic acids research2004
Selection of genomic sequences that bind tightly to Ff gene 5 protein: primer-free genomic SELEX.
Article in Nucleic acids research, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed, 64 citations in OpenAlex.
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- Therapeutic Potential of Aptamer-Protein Interactions.ACS pharmacology & translational science · 2022Review
- Advances in aptamer-based sensing assays for C-reactive protein.Analytical and bioanalytical chemistry · 2022Review
- A New Design for the Fixed Primer Regions in an Oligonucleotide Library for SELEX Aptamer Screening.Frontiers in chemistry · 2020Article
- Application and development of aptamer in cancer: from clinical diagnosis to cancer therapy.Journal of Cancer · 2020Review
- Inside the Black Box: What Makes SELEX Better?Molecules (Basel, Switzerland) · 2019Review
- Updates on Aptamer Research.International journal of molecular sciences · 2019Review
- A new common functional coding variant at the DDC gene change renal enzyme activity and modify renal dopamine function.Scientific reports · 2019Article
- Key Aspects of Nucleic Acid Library Design for in Vitro Selection.International journal of molecular sciences · 2018Review
- Characterization of a cAMP responsive transcription factor, Cmr (Rv1675c), in TB complex mycobacteria reveals overlap with the DosR (DevR) dormancy regulon.Nucleic acids research · 2016Article
- Aptamer-Based Technologies in Foodborne Pathogen Detection.Frontiers in microbiology · 2016Review
- Characterization of the RstB2 protein, the DNA-binding protein of CTXϕ phage from Vibrio cholerae.Virus genes · 2014Article
- An on-bead tailing/ligation approach for sequencing resin-bound RNA libraries.Nucleic acids research · 2012Article
- Challenges and opportunities for small molecule aptamer development.Journal of nucleic acids · 2012Article
- A primer-free method that selects high-affinity single-stranded DNA aptamers using thermostable RNA ligase.Analytical biochemistry · 2011Article
- Genomic SELEX: a discovery tool for genomic aptamers.Methods (San Diego, Calif.) · 2010Review
- Primer-free aptamer selection using a random DNA library.Journal of visualized experiments : JoVE · 2010Article
- Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Single-stranded DNA or RNA libraries used in SELEX experiments usually include primer-annealing sequences for PCR amplification. In genomic SELEX, these fixed sequences may form base pairs with the central genomic fragments and interfere with the binding of target molecules to the genomic sequences. In this study, a method has been developed to circumvent these artificial effects. Primer-annealing sequences are removed from the genomic library before selection with the target protein and are then regenerated to allow amplification of the selected genomic fragments. A key step in the regeneration of primer-annealing sequences is to employ thermal cycles of hybridization-extension, using the sequences from unselected pools as templates. The genomic library was derived from the bacteriophage fd, and the gene 5 protein (g5p) from the phage was used as a target protein. After four rounds of primer-free genomic SELEX, most cloned sequences overlapped at a segment within gene 6 of the viral genome. This sequence segment was pyrimidine-rich and contained no stable secondary structures. Compared with a neighboring genomic fragment, a representative sequence from the family of selected sequences had about 23-fold higher g5p-binding affinity. Results from primer-free genomic SELEX were compared with the results from two other genomic SELEX protocols.
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