Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes

来自 Semantic Scholar

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阅读量：

209

作者：

PR Langer，AA Waldrop，DC Ward

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摘要：

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. Biotinlabeled 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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关键词：

Biochemistry Nucleotide Analog DNA and RNA Polymerase Avidin-Sepharose Antibiotin Antibody Immunoprecipitation

DOI：

10.1073/pnas.78.11.6633

被引量：

2561

年份：

1981