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Expanding the genetic code of a mouse
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 박찬배 | - |
| dc.contributor.author | Lee, Soonjang | - |
| dc.date.accessioned | 2019-10-21T07:25:13Z | - |
| dc.date.available | 2019-10-21T07:25:13Z | - |
| dc.date.issued | 2015-08 | - |
| dc.identifier.other | 20382 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/18684 | - |
| dc.description | 학위논문(석사)--아주대학교 일반대학원 :의생명과학과,2015. 8 | - |
| dc.description.tableofcontents | I.INTRODUCTION 1 II.MATERIALS AND METHOD 3 A. Generation of transgenic mice 3 B. DNA extraction from mouse tail and genotyping PCR 3 C. Isolation of primary mouse embryo fibroblast and SV40 immortalization 3 D. siRNA Transfection 4 E. Cryosection / Confocal microscope 4 III.RESULTS 5 A. Generation of AcKRS.GFP mouse 5 B. Conformation of transgenic mouse 8 C. No dectectable GFP induced by AcK 9 D. Inhibition of Nonsense-mediated mRNA decay through UPF2 Knock down 12 E. Detection of GFP signal in AcKRS.GFP mouse by cryosection 15 IV.DISCUSSION 17 V. CONCLUSION 18 REFERENCES 19 국문요약 21 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | Expanding the genetic code of a mouse | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | Soonjang Lee | - |
| dc.contributor.department | 일반대학원 의생명과학과 | - |
| dc.date.awarded | 2015. 8 | - |
| dc.description.degree | Master | - |
| dc.identifier.localId | 705422 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000020382 | - |
| dc.subject.keyword | aminoacyl-tRNA synthetase | - |
| dc.subject.keyword | Acetyllysine | - |
| dc.subject.keyword | pyrrolysyl-tRNA | - |
| dc.subject.keyword | nonsense-mediated mRNA decay | - |
| dc.description.alternativeAbstract | Genetic code expansion has used the site-specific insertion of unnatural amino acids into proteins(Greiss and Chin, 2011). An aminoacyl-tRNA synthetase and a tRNA are used to specifically insert the unnatural amino acid during mRNA translation, in response to an amber stop codon (UAG) placed at a user-defined site in a gene interest (Davis and Chin, 2012) In this study, I used Acetyllysine(AcK) as unnatural amino acids, N? -acetyl-lysyl-tRNA synthetase (AcKRS) as AcK-tRNA synthetase, pyrrolysyl-tRNA(PylT) as tRNA from Methanosarcina mazei (Mukai et al., 2008). Amber codon was inserted in GFP that is role of reporter gene. AcKRS aminoacylates PylT , and mRNA encoding the full-length GFP bearing an amber codon that directs amino acid incopration. I created AcKRS, GFP mouse, and generated immortalized MEF(mouse embryonic fibroblast). Immortalized MEF(AcKRS.GPF) was treated by AcK., but GFP signal was not detected. I thought that no dectable GFP signal was likely due to the three reasons : First, AcK did not internalize into the system. Second, AcK can be degraded by deacetylase. Third, mRNA was effected by NMD(nonsense-mediated mRNA decay) that can break mRNA containing amber condons. I find the reason that low GFP expression was due to the degradation of mRNA through NMD To investigate GFP signal in AcKRS.GFP mouse I did cryosection and observed by confocal microsope. Because NMD efficiency is various according to organs, I got GFP expression in stomach and muscle in AcKRS.GFP mouse. | - |
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- Graduate School of Ajou University > Department of Biomedical Sciences > 3. Theses(Master)
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