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Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes.
Open accessAbstract
Analogs of dUTP and UTP that contain a biotin molecule covalently bound to the C-5 position of the pyrimidine ring through an allylamine linker arm have been synthesized. These biotin-labeled nucleotides are efficient substrates for a variety of DNA and RNA polymerases in vitro. Polynucleotides containing low levels of biotin substitution (50 molecules or fewer per kilobase) have denaturation, reassociation, and hybridization characteristics similar to those of unsubstituted controls. Biotin-labeled polynucleotides, both single and double-stranded, are selectively and quantitatively retained on avidin-Sepharose, even after extensive washing with 8 M urea, 6 M guanidine hydrochloride, or 99% formamide. In addition, biotin-labeled polynucleotides can be selectively immunoprecipitated in the presence of antibiotin antibody and Staphylococcus aureus protein A. The unique features of biotin-labeled polynucleotides suggest that they will be useful affinity probes for the detection and isolation of specific DNA and RNA sequences.
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Cited by
Hemann, M. T., Strong, M. A., Hao, L.-Y., & Greider, C. W. (2001) · Cell · doi:10.1016/s0092-8674(01)00504-9 · Cited by 1,219
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Prober, J. M., Trainor, G. L., Dam, R. J., Hobbs, F. W., Robertson, C. W., Zagursky, R. J., Cocuzza, A. J., Jensen, M. A., & Baumeister, K. (1987) · Science · doi:10.1126/science.2443975 · Cited by 855
Applications of Aptamers as Sensors
Cho, E. J., Lee, J.-W., & Ellington, A. D. (2009) · Annual Review of Analytical Chemistry · doi:10.1146/annurev.anchem.1.031207.112851 · Cited by 788
RNA Polymerase II Targets Pre-mRNA Splicing Factors to Transcription Sites In Vivo
Misteli, T., & Spector, D. L. (1999) · Molecular Cell · doi:10.1016/s1097-2765(01)80002-2 · Cited by 326
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Metadata
- Authors
- Langer, P. R., Waldrop, A. A., & Ward, D. C.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1981-11
- Volume
- 78
- Issue
- 11
- Pages
- 6633-6637
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2014038908
- Cited by
- 1,145
- References
- 0
- FWCI
- 9.31
- Type
- journal article
- Language
- en
- Open access
- green
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- Langer, P. R., Waldrop, A. A., & Ward, D. C. (1981). Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.78.11.6633
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