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Studies on DEMETERs Promoter and N-terminal Domain Function in Arabidopsis thaliana

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dc.contributor.advisor최연희-
dc.contributor.author조브랜던-
dc.date.accessioned2019-05-07T04:30:57Z-
dc.date.available2019-05-07T04:30:57Z-
dc.date.issued2019-02-
dc.identifier.other000000155419-
dc.identifier.urihttps://hdl.handle.net/10371/151581-
dc.description학위논문 (석사)-- 서울대학교 대학원 : 자연과학대학 생명과학부, 2019. 2. 최연희.-
dc.description.abstractThe development of the female gametophyte in plants is tightly regulated by epigenetic mechanisms to ensure the proper development of seeds. DNA methylation is an evolutionarily conserved mechanism that controls numerous biological processes such as gene imprinting, gene expression, and paramutation. In plants, DNA methylation occurs in the CG, CHG, and CHH contexts. The regulation of DNA methylation is done by a family of DNA demethylase genes including DEMETER (DME), REPRESSOR OF SILENCING 1 (ROS1), DEMETER LIKE 2 (DML2), and DEMETER LIKE 3 (DML3). Both DME and ROS1 are bifunctional DNA glycosylases that initiate active DNA demethylation through the base excision repair pathway. dme mutant plants exhibit severe seed abortion phenotypes as maternally expressed genes including MEDEA (MEA) cannot be activated. The structure of DME includes two splice variants, Spain and Berkeley DME with Spain being 258 amino acids longer on the N-terminal side. DME can further be split up into the N-terminal Domain and the C-terminal domain consisting of the A, G and B domains. Currently, the function of the N-terminal domain is unclear as the C-terminal side was shown to have glycosylase functions in vitro. Dominant negative strategies were done to provide a possible explanation to the function of the N-terminal domain. Transcriptional controls of DME show unexpected expression patterns opening the possibility that another factor controls DME expression. A series of differential DME fragments also showed unexpected GFP expression patterns.-
dc.description.tableofcontentsABSTRACT. 1
CONTENTS. 3
LIST OF FIGURES 5
LIST OF TABLES 7
ABBREVIATIONS. 8
I. INTRODUCTION. 9
1. Overview of Plant Reproductive Development 9
2. Epigenetic Control for Proper Seed Development in Plants 13
3. Mechanism of Active DNA Demethylation in Plants 14
4. Role of DEMETER, a DNA glycosylase, in plant development. 16
5. Structural Analysis of DEMETER. 19
6. Purpose of this study. 21
II. MATERIALS AND METHODS 23
1. Plant material and growth conditions. 23
2. Recombinant plasmid construction. 23
3. Agrobacterium tumefaciens transformation and plant transformation by floral dipping. 24
4. Yeast 2 Hybrid protocol 25
5. Microscopy for GFP Analysis 26
III. RESULTS. 31
1. Investigations of the possible binding partners of DEMETERs N-Terminal Domain (NTD) 31
1.1 DEMETERs Interaction with and Ubiquitinligase (E3). 31
1.2 DEMETERs Self-Interaction. 33
1.3 DEMETERs N-terminal cDNA Overexpression. 37
1.4 DEMETERs A-Domain Effect on Expression Patterns 40
1.5 DEMETER cDNA Ectopic Expression. 42
2. Analysis of the relationship between DEMETERs Promoter and its N-terminal Domain in expression patterns 45
IV. DISCUSSION. 49
V. REFERENCES 52
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dc.language.isoeng-
dc.publisher서울대학교 대학원-
dc.subject.ddc570-
dc.titleStudies on DEMETERs Promoter
and N-terminal Domain Function in Arabidopsis thaliana
-
dc.typeThesis-
dc.typeDissertation-
dc.contributor.AlternativeAuthorStudies on DEMETER's Promoter and N-terminal Domain Function in Arabidopsis thaliana-
dc.description.degreeMaster-
dc.contributor.affiliation자연과학대학 생명과학부-
dc.date.awarded2019-02-
dc.identifier.uciI804:11032-000000155419-
dc.identifier.holdings000000000026▲000000000039▲000000155419▲-
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