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Structural and functional study of Streptococcus pyogenes Csn2, CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-associated protein

DC Field Value Language
dc.contributor.advisor배의영-
dc.contributor.author구윤-
dc.date.accessioned2017-07-14T06:38:11Z-
dc.date.available2017-07-14T06:38:11Z-
dc.date.issued2012-08-
dc.identifier.other000000003257-
dc.identifier.urihttps://hdl.handle.net/10371/125790-
dc.description학위논문 (석사)-- 서울대학교 대학원 : 농생명공학부, 2012. 8. 배의영.-
dc.description.abstractThe CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR-associated proteins) system functions as an adaptive immune system that protects prokaryotic cells from invading genomic nucleic acids, such as bacteriophages and conjugative plasmids.
Csn2 is a Nmeni subtype-specific Cas protein, recently classified as type Ⅱ–A, required for new spacer acquisition in the adaptation process of this immune system. In this study, the crystal structure of Streptococcus pyogenes Csn2 revealed a ring-like quaternary structure including calcium binding sites. Also, the Csn2 has a non-specific double-stranded (ds-) DNA-binding activity.
In summary, the results support the suggestions of Csn2 functioning at the CRISPR adaptation stage.
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dc.description.tableofcontentsAbstract ⅰ
Contents ⅲ
List of Figures ⅳ
List of Tables ⅴ
List of Abbreviations ⅵ

1. INTRODUCTION 1
2. MATERIALS AND METHODS 4
2.1 Protein expression and purification 4
2.2 Expression and purification selenomethionyl protein 5
2.3 Mass spectrometry 6
2.4 Crystallization 6
2.5 Structure determination and refinement 7
2.6 Metal analysis 8
2.7 Oligomeric state analysis 8
2.8 Electrophoretic mobility shift assay 9
3. RESULTS 11
3.1 Expression, purification and crystallization of Csn2 11
3.2 Overall structure of S. pyogenes Csn2 15
3.3 Calcium binding sites in Csn2 protomer 25
3.4 S. pyogenes Csn2 binds to ds-DNA 26
4. DISCUSSION 32
5. ACCESSION NUMBERS 35
6. REFERENCES 36


Abstract in Korean 42
Acknowledgement 44
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dc.formatapplication/pdf-
dc.format.extent1628692 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 대학원-
dc.subjectCRISPR-Cas system-
dc.subjectStreptococcus pyogenes SF370-
dc.subjectCsn2-
dc.subjectx-ray crystallography-
dc.subjectDNA binding-
dc.subject.ddc630-
dc.titleStructural and functional study of Streptococcus pyogenes Csn2, CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-associated protein-
dc.typeThesis-
dc.description.degreeMaster-
dc.citation.pagesⅵ, 44-
dc.contributor.affiliation농업생명과학대학 농생명공학부-
dc.date.awarded2012-08-
Appears in Collections:
College of Agriculture and Life Sciences (농업생명과학대학)Dept. of Agricultural Biotechnology (농생명공학부)Theses (Master's Degree_농생명공학부)
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