DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | Lee, Young-Hoon | - |
dc.contributor.advisor | 이영훈 | - |
dc.contributor.author | Kim, Mee-Hyein | - |
dc.contributor.author | 김미현 | - |
dc.date.accessioned | 2011-12-13T05:00:14Z | - |
dc.date.available | 2011-12-13T05:00:14Z | - |
dc.date.issued | 1995 | - |
dc.identifier.uri | http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=98735&flag=dissertation | - |
dc.identifier.uri | http://hdl.handle.net/10203/32691 | - |
dc.description | 학위논문(석사) - 한국과학기술원 : 화학과, 1995.2, [ vi, 58 p. ] | - |
dc.description.abstract | Aptamers 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.language | eng | - |
dc.publisher | 한국과학기술원 | - |
dc.title | In vitro selection of DNA aptamers binding to interleukin-6 | - |
dc.title.alternative | Interleukin-6에 결합하는 DNA aptamer의 선별 | - |
dc.type | Thesis(Master) | - |
dc.identifier.CNRN | 98735/325007 | - |
dc.description.department | 한국과학기술원 : 화학과, | - |
dc.identifier.uid | 000933053 | - |
dc.contributor.localauthor | Lee, Young-Hoon | - |
dc.contributor.localauthor | 이영훈 | - |
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