Characterization of Endospore Proteins Associated with DNA Protection
- Title
- Characterization of Endospore Proteins Associated with DNA Protection
- Alternative Title
- 디옥시리보핵산 보호와 관련된 내생포자 단백질의 특성 분석
- Author(s)
- Hyogyung Shin
- DGIST Authors
- Hyogyung Shin ; Yoori Kim ; Young-Sam Lee
- Advisor
- 김유리
- Co-Advisor(s)
- Young-Sam Lee
- Issued Date
- 2023
- Awarded Date
- 2023-08-01
- Type
- Thesis
- Description
- Clostridium perfringens; endospore; small acid-soluble protein; DNA protection; sortase-mediated fluorescent labeling
- Table Of Contents
-
Ⅰ. Introduction 1
Ⅱ. Materials and Methods 9
2.1 Wild-type Ssp4 expression plasmids 9
2.2 Purification of Ssp4 9
2.3 Preparation of DNA 10
2.4 Electrophoretic mobility shift assay (EMSA) 11
2.5 DNA protection assay against UV irradiation 12
2.6 DNA protection assay against DNase I degradation 13
2.7 Site-directed mutagenesis for N-terminal GGG-Ssp4 14
2.8 Purification of sortase A 15
Ⅲ. Results 17
3.1 Purification of Ssp4:D36 and Ssp4:G36 proteins 17
3.2 DNA binding affinity for Ssp4 variants for random-sequence DNA 20
3.3 DNA binding affinity for Ssp4 variants for AT-rich DNA 24
3.4 DNA protection activity of Ssp4 variants 27
3.5 Sortase-mediated fluorescent labeling 31
3.5.1 Construction of N-terminal GGG-Ssp4 vector 31
3.5.2 Purification of sortase A 34
3.5.3 Establishment of sortase-mediated fluorescent labeling strategy for Ssp4 36
Ⅳ. Discussion 40
References 44
Summary in Korean 56
- URI
-
http://hdl.handle.net/20.500.11750/46423
http://dgist.dcollection.net/common/orgView/200000686904
- Degree
- Master
- Department
- Department of New Biology
- Publisher
- DGIST
- Related Researcher
-
-
Kim, Yoori
- Research Interests Biophysics; Biochemistry
-
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- Appears in Collections:
- Department of New Biology Theses Master
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