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Expression and purification of JAZ3 protein and a transcriptional factor MYC2 from Arabidopsis thaliana : 애기장대 유래 전사조절인자 JAZ3 와 전사인자 MYC2 단백질 복합체 발현 및 정제
DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | 이상기 | - |
dc.contributor.author | 김두원 | - |
dc.date.accessioned | 2017-07-14T06:44:25Z | - |
dc.date.available | 2017-07-14T06:44:25Z | - |
dc.date.issued | 2015-08 | - |
dc.identifier.other | 000000049848 | - |
dc.identifier.uri | https://hdl.handle.net/10371/125905 | - |
dc.description | 학위논문 (석사)-- 서울대학교 대학원 : 농생명공학부(응용생명화학전공), 2015. 6. 이상기. | - |
dc.description.abstract | Jasmonate is a plant hormone that regulates development, growth and wound response. There are many jasmonate derivatives in nature. Particularly, jasmonyl isoleucine serves as a bioactive molecule in the jasmonate signaling pathway. In fact, the jasmonate signaling is regulated by interactions among four different proteins, including MYC2, JAZ, NINJA, and TOPLESS. Among them, MYC2 and JAZ have critical role in the jasmonate signaling pathway. JAZ binds to MYC2 and represses the jasmonate signaling. Especially, intermolecular interaction of JAZ and MYC2 is attributable to JID motif of MYC2 and Jas motif of JAZ. Nonetheless these interactions play a crucial role in the jasmonate signaling pathway, molecular details of JAZ and MYC2 remain unknown, mainly due to the insoluble properties of each JAZ and MYC2 protein in vitro. In this thesis, I report various approaches that could yield a protein crystal of Jas and JID motif in JAZ and MYC2, respectively. Eight fusion proteins were produced. In those fused proteins, two different constructs for Jas motif and four for JID region are linked using Tobacco Etch Virus (TEV) protease cleavage sequence. Various chromatographic analysis indicated that JAZ forms a complex with MYC2 by cleaving the TEV protease cleavage site. Although crystallization of the complex has not been achieved at this moment, this study provides valuable information for producing the JAZ-MYC2 complex in vitro. | - |
dc.description.tableofcontents | ABSTRACT .............................................................................................. i
CONTENTS ........................................................................................... iii LIST OF FIGURES ................................................................................ iv LIST OF TABLES................................................................................... v LIST OF ABBRIVIATION ..................................................................... vi INTRODUCTION ....................................................................................1 MATERIALS AND METHODS Cloning of JAZ3-TEV-MYC2 fusion proteins ................................5 Evaluation of Expression level and solubility of JAZ3-TEV-MYC2 fusion proteins ...............................................................................5 Expression and purification of JAZ3-TEV-MYC2 fusion proteins and mutants of J352-TEV-M324 ...............................................7 Methylation of J352-TEV-M324 ....................................................8 Proteolysis of J352-TEV-M324 and methylated J352-TEV-M324 with TEV .............................................................................8 Crystallization .....................................................................................9 RESULTS Expression and purification of JAZ3-TEV-MYC2 fusion proteins and mutants ................................................................................. 10 DISCUSSION ........................................................................................ 13 REFERENCES ...................................................................................... 15 | - |
dc.format | application/pdf | - |
dc.format.extent | 1268614 bytes | - |
dc.format.medium | application/pdf | - |
dc.language.iso | en | - |
dc.publisher | 서울대학교 대학원 | - |
dc.subject | jasmonate | - |
dc.subject | jasmonyl isoleucine | - |
dc.subject | JA signaling | - |
dc.subject | MYC2 | - |
dc.subject | Jasmonate-ZIM domain3 | - |
dc.subject.ddc | 630 | - |
dc.title | Expression and purification of JAZ3 protein and a transcriptional factor MYC2 from Arabidopsis thaliana | - |
dc.title.alternative | 애기장대 유래 전사조절인자 JAZ3 와 전사인자 MYC2 단백질 복합체 발현 및 정제 | - |
dc.type | Thesis | - |
dc.contributor.AlternativeAuthor | Duwon Kim | - |
dc.description.degree | Master | - |
dc.citation.pages | vi, 40 | - |
dc.contributor.affiliation | 농업생명과학대학 농생명공학부 | - |
dc.date.awarded | 2015-08 | - |
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