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Differential Function of the Tail Fiber Proteins and Receptor Binding Protein on Host Specificity of Bacteriophage SCB3 : 박테리오파지 SCB3의 숙주 특이성에 대한 Tail Fiber Protein과 수용체 결합 단백질의 기능 연구

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dc.contributor.advisor유상렬-
dc.contributor.author박부경-
dc.date.accessioned2017-07-14T06:40:45Z-
dc.date.available2017-07-14T06:40:45Z-
dc.date.issued2014-02-
dc.identifier.other000000017371-
dc.identifier.urihttps://hdl.handle.net/10371/125834-
dc.description학위논문 (석사)-- 서울대학교 대학원 : 농생명공학부, 2014. 2. 유상렬.-
dc.description.abstractBacteriophage which can infect specific bacteria has emerged as a new tool to control various food-borne pathogens. A novel bacteriophage SCB3 targeting Salmonella enterica serovar Typhimurium was isolated from chicken skin. Morphological analysis revealed that phage SCB3 belongs to the Siphoviridae family. The genome of phage SCB3 consists of 108,773-bp nucleotides excluding terminal redundancy with 153 open reading frames (ORFs). Additional phylogenetic analysis revealed that phage SCB3 is classified as T5-like phage. Interestingly, phage SCB3 could infect only rough strains which do not have O-antigen of lipopolysaccharides even though BtuB (the vitamin B12 uptake outer membrane protein) is used as a host receptor. Comparative genomic analysis of phage SCB3 with other T5-like phage SPC35 revealed that the specificity to rough strains of phage SCB3 may be associated with tail fiber genes. Swapping of the tail fiber gene of phage SCB3 with that of phage SPC35 enabled phage SCB3 to infect wild type Salmonella, suggesting that the tail fiber protein is involved in the interaction of phage with O-antigen. These results indicate that phage tail fiber protein may play an ancillary role in infection of bacteria by phages using BtuB as a receptor. Therefore, it experimentally supports that tail fiber protein of phage SCB3 is one of the major determinant for host specificity, along with the receptor binding protein.-
dc.description.tableofcontentsABSTRACT................................................................................................................i
CONTENTS.............................................................................................................iii
I. INTRODUCTION...............................................................................................1
II. MATERIALS AND METHODS......................................................................5
1. Bacterial strains and growth condition.........................................................5
2. Bacteriophage isolation and propagation…..…………..........................5
3. Transmission electron microscopy….....................................................7
4. Spotting assay and efficiency of plating…………..…………………..8
5. Construction of deletion mutants and complementation……………8
6. One step growth curve…………………………………………...........9
7. Bioinformatics analysis.……………….........................................10
8. Phage DNA extraction…………………………………………..……..11
9. Construction of the mutant phages…………………..............................12
10. Adsorption assay………………………………………...……...............12
11. Bacterial challenge assay and phage challenge assay…............………13
12. Beta-galactosidase assay and gtrABC1 overexpression…....…..…14
III. RESULTS.................................................................................................20
1. Isolation of SCB3 and its morphology………………..….…......…….20
2. Host range of SCB3……………………………..……...…........……..23
3. Receptor determination of SCB3…..........................................................26
4. One step growth curve of SCB3…………...............................................33
5. Whole genome sequence of SCB3………..……........................................35
6. Comparative genomic analysis with SPC35……………….…….….....38
7. Swapping the tail fiber genes of SCB3 with those of SPC35……..43
8. Resistant mutants of LT2(c) to SCB3 TFM infection occurred by glucosylating their O-antigen.…………...…………………..……51
IV. DISCUSSION.........................................................................................57
V. REFFERENCES....................................................................................64
국문초록.......................................................................................................71
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dc.formatapplication/pdf-
dc.format.extent3299697 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 대학원-
dc.subjectSalmonella Typhimurium-
dc.subjectBacteriophage-
dc.subjectTail fiber protein-
dc.subjectReceptor binding protein-
dc.subjectHost specificity-
dc.subject.ddc630-
dc.titleDifferential Function of the Tail Fiber Proteins and Receptor Binding Protein on Host Specificity of Bacteriophage SCB3-
dc.title.alternative박테리오파지 SCB3의 숙주 특이성에 대한 Tail Fiber Protein과 수용체 결합 단백질의 기능 연구-
dc.typeThesis-
dc.contributor.AlternativeAuthorBookyung Park-
dc.description.degreeMaster-
dc.citation.pagesiv, 72-
dc.contributor.affiliation농업생명과학대학 농생명공학부-
dc.date.awarded2014-02-
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