Bio-manufacturing logical bio-circuits and thereof mounted bio-systems for diagnosing and indexing severity of Inflammatory Bowel Disease
염증성 장질환 진단 및 질환 심각도 인덱싱 가능한 논리 생체회로와 이를 탑재한 바이오-시스템 제조
- 주제(키워드) IBD , Living therapeutics , Gut barrier dysfuction , Inflammatory biomarker , layered logic gate , Disease Severity
- 발행기관 고려대학교 대학원
- 지도교수 박제원
- 발행년도 2025
- 학위수여년월 2025. 2
- 학위명 석사
- 학과 및 전공 대학원 의생명융합과학과
- 세부전공 바이오의과학
- 원문페이지 99 p
- 실제URI http://www.dcollection.net/handler/korea/000000290951
- UCI I804:11009-000000290951
- DOI 10.23186/korea.000000290951.11009.0001890
- 본문언어 영어
목차
TABLE OF CONTENTS
ABSTRACT i
국문 초록 iii
TABLE OF CONTENTS v
LIST OF TABLES vii
LIST OF FIGURES viii
NOMENCLATURE x
CHAPTER 1. INTRODUCTION 1
1.1 Inflammatory Bowel Disease 1
1.1.1 Etiology of Inflammatory Bowel Disease 2
1.2 IBD-diagnosis: sucralose as an indicator of gut barrier dysfuction 4
1.3 IBD-diagnosis: kynurenine as an inflammatory biomarker 8
1.4 Indexing the severity of IBD: heme as an indicator for intestinal hemorrhage 10
1.5 Living bio-therapeutics for IBD-diagnosis 11
1.5.1 Modus Operandi: Gut barrier dysfunction-sensing bio-circuit 12
1.5.2 Modus Operandi: Inflammatory signal-sensing bio-circuit 14
1.5.3 Modus Operandi: Gastrointestinal hemorrhaging-sensing bio-circuit 16
1.6 Aim of thesis 18
CHAPTER 2. MATERIALS AND METHODS 21
2.1 Chemicals and enzymes 21
2.2 Bacterial strains, plasmid, and growth condition 22
2.3 DNA manipulation and sequencing 29
2.4 Knockout of the lacI gene in the chromosome to eliminate circuit noise 30
2.5 Bio-part preparation 32
2.6 PCR amplification 33
2.7 Genetic layered logic gate circuits for addressing multiple biomarkers 37
2.8 Genetic circuit design for IBD sensing 40
2.9 Induction conditions in engineered E. coli strains. 41
2.9.1 Induction conditions for sucralose-sensing bio-circuit 42
2.9.2 Induction conditions for kynurenine-sensing bio-circuit 43
2.9.3 Induction conditions for heme-sensing bio-circuit 44
2.9.4 Induction conditions for the orthogonality test 45
2.9.5 Induction conditions for NOT logic gate bio-circuit 46
2.9.6 Induction conditions for AND logic gate bio-circuit 47
2.9.7 Induction conditions for layered logic gate bio-circuit 48
2.10 Reporter gene assays 49
CHAPTER 3. RESULTS 51
3.1 Expression levels of each biomarker sensing bio-system mounted in E. coli K12 MG1655 (ΔlacI) 51
3.1.1 Evaluation of the sucralose-sensing bio-system 52
3.1.2 Evaluation of the kynurenine-sensing bio-system 54
3.1.3 Evaluation of the heme-sensing bio-system 56
3.1.4 Orthogonality test of each biomarker-sensing bio-system 58
3.2 Expression levels of logic gate bio-system mounted in E. coli K12 MG1655 60
3.2.1 Evaluation of the NOT logic gate bio-system 61
3.2.2 Evaluation of the AND logic gate bio-system 63
3.2.3 Evaluation of the layered logic gate bio-system 67
CHAPTER 4. DISCUSSION AND FUTURE PERSPECTIVES 71
REFERENCES 75
Appendix 81
