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KMID : 0578319950050060555
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Molecules and Cells
1995 Volume.5 No. 6 p.555 ~ p.562
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In Vitro Selection if DNA Aptamers Binding to the Recombinant Human Interleukin-6
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Kim, MeeHyein
Jeoung, Yeon-Hee/Lee, Seong Jae/Choi, Inpyo/Pyun, Kwang-Hp/Lee, Younghonn
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Abstract
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IL-6 is known to inhibit growth and induce differentiation of several myeloid leukemia cell lines. With the recombinant human interleukin-6 (IL-6) as a target protein, DNA aptamers specifically binding to IL-6 were isolated from a random oligodeoxyribonucleotide pool. A large pool of 96-mer oligodeoxyribonucleotides, made up of 60 bases of random sequences flanked by the defined regions of primer binding sites at the 5´ and 3´ ends, was synthesized. This randomly generated oligodeoxyribonucleotide pool was denatured to generate singles-stranded DNA and subjected to in vitro selection with affinity chromatography and in vitro amplification with polymerase chain reaction (PCR) for enrichment in single-stranded DNA aptamers specifically binding to IL-6. Multiple rounds of successive affinity chromatography and PCR resulted in the continued purification of binding species. The gel retardation experiment showed that aptamers had affinity to IL-6 with dissociation constants of the order of magnitude of 10^(-6) M. The sequences of the aptamers were identified. A homology search revealed that they were categorized into at least four groups according to the consensus sequences among them.
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KEYWORD
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