Factors affecting the alternative splicing of canine glutamine synthetase gene개 글루타민 합성효소 유전자의 특이적 스플라이싱에 영향을 주는 factor의 확인

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dc.contributor.advisorPark, Chan-Kyu-
dc.contributor.advisor박찬규-
dc.contributor.authorLee, Yoon-Jung-
dc.contributor.author이윤정-
dc.date.accessioned2011-12-12T08:53:56Z-
dc.date.available2011-12-12T08:53:56Z-
dc.date.issued2004-
dc.identifier.urihttp://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=237818&flag=dissertation-
dc.identifier.urihttp://hdl.handle.net/10203/28025-
dc.description학위논문(석사) - 한국과학기술원 : 생명과학과, 2004.2, [ iv, 45 p. ]-
dc.description.abstractGlutamine synthetase (GS) catalyzes the ATP-dependent conversion of ammonia and glutamate to glutamine. In the previous study, we found an additional exon (120 bp) in the first intron of GS gene. By means of alternative splicing, the GS gene produces an altered form of GS transcript with 5`-untranslated region (UTR) containing the additional exon. This alternative transcript is abundantly expressed in brain, whereas it is found at lower levels in other tissues. We have performed an analysis on the cis-acting regulatory sequences that reside in the intron immediately upstream of this mini-exon, designated here as intron 1a, using the exon-trapping system. We constructed a series of mutants, in which either whole or partial GS intron 1a was replaced with GS intron 4 which is efficiently spliced out. When this constructs were transfected into MDCK and Neuro-2A cells, we found that both the branch-site and the polypyrimidine tract in GS intron 1a serve as cis-acting elements in brain-specific alternative splicing. Furthermore, we observed that a sequence motif in the new exon has no SRp40 binding sequence in near 3``(acceptor) exon-intron junction. So we suspected that SRp40 functions as exonic trans-acting factor in the brain-specific splicing of canine GS gene. But in the exon-trapping experiment with the exonic mutant minigene constructs, it seems that the absence of SRp40 binding sequence motif in exon 1`` is not related to GS alternative splicing. On the other hand, we describe a novel method for identifying candidate trans-acting factors that are involved in regulating GS alternative splicing. We generated a cell line that stably expresses a GS minigene-based GFP reporter construct, such that the levels of green-fluorescent protein (GFP) expressed in the cell reflect the splicing state of the GS gene.eng
dc.languageeng-
dc.publisher한국과학기술원-
dc.subjectGLUTAMINE SYNTHETASE GENE-
dc.subjectALTERNATIVE SPLICING-
dc.subject특이적 스플라이싱-
dc.subject글루타민 합성효소 유전자-
dc.titleFactors affecting the alternative splicing of canine glutamine synthetase gene-
dc.title.alternative개 글루타민 합성효소 유전자의 특이적 스플라이싱에 영향을 주는 factor의 확인-
dc.typeThesis(Master)-
dc.identifier.CNRN237818/325007 -
dc.description.department한국과학기술원 : 생명과학과, -
dc.identifier.uid020023457-
dc.contributor.localauthorPark, Chan-Kyu-
dc.contributor.localauthor박찬규-
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