Sortase a engineering in escherichia coli using tripartite split green fluorescent protein system세 조각 스플릿 녹색형광단백질 시스템을 이용한 대장균에서의 전이효소 솔테이즈 에이 개령

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dc.contributor.advisorJeong, Ki Jun-
dc.contributor.advisor정기준-
dc.contributor.authorLee, Roojin-
dc.contributor.author이루진-
dc.date.accessioned2017-03-29T02:34:36Z-
dc.date.available2017-03-29T02:34:36Z-
dc.date.issued2016-
dc.identifier.urihttp://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=649482&flag=dissertationen_US
dc.identifier.urihttp://hdl.handle.net/10203/221529-
dc.description학위논문(석사) - 한국과학기술원 : 생명화학공학과, 2016.2 ,[v, 42 p. :]-
dc.description.abstractStaphylococcus aureus Sortase A, the trans-peptidase which ligate the protein to the gram-positive cell wall is widely used for the post-translational protein ligation. It has been highlighted due to its site-specific catalytic activity with short target motif. However Staphylococcus aureus sortase A is highly depends on the existence of $Ca^{2+}$ ion, therefore it is hard to utilize sortase A in vivo. To over-come this problem, I have developed high throughput screening system to isolate $Ca^{2+}$ independent SrtA mutant in Escherichia coli based on the tripartite split green fluorescent protein (GFP) system. Reassembly of tripartite split GFP catalyzed by $Ca^{2+}$ independent sortase A could be detected by high speed fluorescence activated cell sorter (FACS). After several round sorting, I found and characterized $Ca^{2+}$ independent sortase A mutant which had different mutation point with rationally engineered sort-ase A. The catalytic activity also validated using in vitro and in vivo analysis with several types of sub-strate. This research is not only an easy system of enzyme engineering, but also the first engineering application of tripartite split GFP system.-
dc.languageeng-
dc.publisher한국과학기술원-
dc.subjectSortase A-
dc.subjectTripartite split green fluorescent protein system-
dc.subjectHigh-throughput screening-
dc.subjectEnzyme engineering-
dc.subjectEscherichia coli-
dc.subjectStaphylococcus aureus-
dc.subject솔테이즈 에이-
dc.subject세 조각 스플릿 녹색형광단백질-
dc.subject고효율 발굴 시스템-
dc.subject효소 개량-
dc.subject무작위 돌연변이-
dc.titleSortase a engineering in escherichia coli using tripartite split green fluorescent protein system-
dc.title.alternative세 조각 스플릿 녹색형광단백질 시스템을 이용한 대장균에서의 전이효소 솔테이즈 에이 개령-
dc.typeThesis(Master)-
dc.identifier.CNRN325007-
dc.description.department한국과학기술원 :생명화학공학과,-
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