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Genome-wide transcriptome analysis of Enterohemorrhagic Escherichia coli (EHEC) isolates for identification of fresh produces-specific virulence factors
식물성 식품 유래 장출혈성 대장균의 전사체 분석 및 식물성 식품 기반 특이적 유해 인자의 특성 규명

DC Field Value Language
dc.contributor.advisor유상렬-
dc.contributor.author김연경-
dc.date.accessioned2017-07-14T06:45:23Z-
dc.date.available2017-07-14T06:45:23Z-
dc.date.issued2016-02-
dc.identifier.other000000131824-
dc.identifier.urihttps://hdl.handle.net/10371/125924-
dc.description학위논문 (석사)-- 서울대학교 대학원 : 농생명공학부 농생명공학전공, 2016. 2. 유상렬.-
dc.description.abstractEnterohemorrhagic Escherichia coli (EHEC) is a food-borne pathogen causing serious threats to human. Recent studies have revealed that many outbreaks of EHEC were mediated by spoiled produce such as raw sprouts or lettuce. To investigate how the plant-derived EHEC cells adapt to fresh produces, EHEC #736 isolated from kimchi in Korea was cultivated with canola spouts (Brassica napus) and water dropwort (Oenanthe javanica). Planktonic bacteria in fresh produces were subjected to RNA sequencing to achieve bacterial mRNA profiles responding to contacts with canola sprouts and water dropwort. As a result, among a total of 4,882 annotated coding sequences, 6.06% and 5.12% of total genes were significantly up- or down-regulated when exposed to canola sprouts and water dropwort respectively after 1 hour. Similarly, 5.90% and 4.53% of total genes were significantly up- or down-regulated when exposed to canola sprouts and water dropwort respectively for 3 hours. Genes with different expression (p<0.05) could be grouped mainly into five categories: translation and ribosome biogenesis, cell motility, carbohydrate transport, energy conversion and secondary metabolites biosynthesis/transport. In detail, multiple nutrition transport systems for carbohydrate, amino acid and nitrate that are known to be prevalent in plants were activated in EHEC #736. This indicates the availability to EHEC #736 of a variety of factors released from fresh produces that may promote the growth of bacteria. Particularly, RNA-Seq revealed that iron uptake related genes including Fur (Ferric-uptake regulator) regulons are differentially expressed when the bacteria were exposed to plants. It was further demonstrated that EHEC #736 ΔentB mutant defective in siderophore biosynthesis had growth defects in water dropwort, and the defective phenotype was complemented by the addition of ferrous ion. This indicates that iron uptake is essential for survival of EHEC in plant. Considering the importance of iron acquisition for bacterial survival, iron disrupting mechanism, such as treating gallium, a transition metal, might prevent fresh produce from bacterial contamination.-
dc.description.tableofcontentsI. INTRODUCTION 6
II. MATERIALS AND METHODS 10
1. Preparation of canola sprouts and water dropwort 10
2. Bacterial strains and growth condition 10
3. Construction of bacterial deletion mutants 11
4. In planta assay 12
5. RNA extraction and sequencing 13
6. Quantitative real-time PCR (qRT-PCR) analysis 15
7. Heat map generation 16
8. Biofilm reduction assay 16
III. RESULTS 21
1. Growth of EHEC #736 in canola sprouts and water dropwort 21
2. Transcriptional response of EHEC #736 to canola sprouts and water dropwort 24
3. Expression patterns of genes with known function 36
4. qRT-PCR for DEGs of EHEC #736 exposed to canola sprouts and water dropwort 46
5. Transcriptional response of genes related to iron metabolism 48
6. Enterobactin is required for EHEC #736 survival in water dropwort 52
7. Gallium disrupts iron metabolism of EHEC #736 on water dropwort and has antibiofilm activity 56
IV. DISCUSSION 59
V. CONCLUSION 66
VI. REFFERENCES 67
국문초록 76
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dc.formatapplication/pdf-
dc.format.extent1525773 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 대학원-
dc.subjectEHEC-
dc.subjectfresh produce-
dc.subjecttranscriptome analysis-
dc.subjectRNA-Seq-
dc.subjectiron acquisition-
dc.subjectgallium-
dc.subject.ddc630-
dc.titleGenome-wide transcriptome analysis of Enterohemorrhagic Escherichia coli (EHEC) isolates for identification of fresh produces-specific virulence factors-
dc.title.alternative식물성 식품 유래 장출혈성 대장균의 전사체 분석 및 식물성 식품 기반 특이적 유해 인자의 특성 규명-
dc.typeThesis-
dc.contributor.AlternativeAuthorYeonkyung Kim-
dc.description.degreeMaster-
dc.citation.pages78-
dc.contributor.affiliation농업생명과학대학 농생명공학부-
dc.date.awarded2016-02-
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College of Agriculture and Life Sciences (농업생명과학대학)Dept. of Agricultural Biotechnology (농생명공학부)Theses (Master's Degree_농생명공학부)
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