In vitro selection of DNA aptamers binding to interleukin-6Interleukin-6에 결합하는 DNA aptamer의 선별

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dc.contributor.advisorLee, Young-Hoon-
dc.contributor.advisor이영훈-
dc.contributor.authorKim, Mee-Hyein-
dc.contributor.author김미현-
dc.date.accessioned2011-12-13T05:00:14Z-
dc.date.available2011-12-13T05:00:14Z-
dc.date.issued1995-
dc.identifier.urihttp://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=98735&flag=dissertation-
dc.identifier.urihttp://hdl.handle.net/10203/32691-
dc.description학위논문(석사) - 한국과학기술원 : 화학과, 1995.2, [ vi, 58 p. ]-
dc.description.abstractAptamers are DNA or RNA molecules that bind specific molecular targets. With the recombinant human interleukin-6 (IL-6) as a target protein, which has been known to inhibit growth and induce differentiation of several myeloid leukemia cell lines, a set of DNA aptamers binding to IL-6 from a random oligodeoxyribonucleotide pool was isolated. A large pool of 96-mer oligodeoxyribonucleotides was chemically synthesized that was made up of 60 bases of random sequences flanked by the defined regions of primer binding sites at the $5^\prime$ and $3^\prime$ ends. This randomly generated oligodeoxyribonucleotide pool renaturated at $25\,^\circ\!C$ for proper folding of single-stranded DNAs after denatured at $95\,^\circ\!C$ was subjected to in vitro selection with affinity chromatography and in vitro amplification with polymerase chain reaction (PCR) for enrichment in DNA aptamers specifically binding to IL-6. Multiple rounds of affinity chromatography and PCR resulted in the continued purification of binding species. The sequences of the aptamers and their binding properties for IL-6 will be determined. Since a binding strand can not be chosen from the sequencing data, binding affinity of labeled two single-stranded DNAs was tested by gel retardation assay, respectively. The gel retardation experiment showed that aptamers had affinity to IL-6 and their dissociation constants were about $\sim10^{-6}$ M. A homology search revealed that they were catalogued into at least three groups according to the consensus sequences among them. The consensus sequences are likely to participate in forming IL-6 binding motifs.eng
dc.languageeng-
dc.publisher한국과학기술원-
dc.titleIn vitro selection of DNA aptamers binding to interleukin-6-
dc.title.alternativeInterleukin-6에 결합하는 DNA aptamer의 선별-
dc.typeThesis(Master)-
dc.identifier.CNRN98735/325007-
dc.description.department한국과학기술원 : 화학과, -
dc.identifier.uid000933053-
dc.contributor.localauthorLee, Young-Hoon-
dc.contributor.localauthor이영훈-
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CH-Theses_Master(석사논문)
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