DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | Jeong, Ki Jun | - |
dc.contributor.advisor | 정기준 | - |
dc.contributor.author | Lee, Roojin | - |
dc.contributor.author | 이루진 | - |
dc.date.accessioned | 2017-03-29T02:34:36Z | - |
dc.date.available | 2017-03-29T02:34:36Z | - |
dc.date.issued | 2016 | - |
dc.identifier.uri | http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=649482&flag=dissertation | en_US |
dc.identifier.uri | http://hdl.handle.net/10203/221529 | - |
dc.description | 학위논문(석사) - 한국과학기술원 : 생명화학공학과, 2016.2 ,[v, 42 p. :] | - |
dc.description.abstract | Staphylococcus 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.language | eng | - |
dc.publisher | 한국과학기술원 | - |
dc.subject | Sortase A | - |
dc.subject | Tripartite split green fluorescent protein system | - |
dc.subject | High-throughput screening | - |
dc.subject | Enzyme engineering | - |
dc.subject | Escherichia coli | - |
dc.subject | Staphylococcus aureus | - |
dc.subject | 솔테이즈 에이 | - |
dc.subject | 세 조각 스플릿 녹색형광단백질 | - |
dc.subject | 고효율 발굴 시스템 | - |
dc.subject | 효소 개량 | - |
dc.subject | 무작위 돌연변이 | - |
dc.title | Sortase a engineering in escherichia coli using tripartite split green fluorescent protein system | - |
dc.title.alternative | 세 조각 스플릿 녹색형광단백질 시스템을 이용한 대장균에서의 전이효소 솔테이즈 에이 개령 | - |
dc.type | Thesis(Master) | - |
dc.identifier.CNRN | 325007 | - |
dc.description.department | 한국과학기술원 :생명화학공학과, | - |
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