표제지

목차

1. Endoplasmic Reticulum-Located PDAT1- 2 from Castor Bean Enhances Hydroxy Fatty Acid Accumulation in Transgenic Plants (Plant Cell Physiology) / Hyun Uk Kim ; Kyeong-Ryeol Lee ; Young sam Go ; Jin Hee Jung ; Mi-Chung Suh ; Jong Bum Kim 4

Introduction 4

Results 5

The existence of three PDATs and PDAT1-2 is specific to the castor bean plant 5

Expression of castor bean PDAT 6

ER localization of castor bean PDAT1-1 and PDAT1-2 6

Castor bean PDAT1-2 boosts accumulation of hydroxy fatty acids in seeds 6

Castor bean PDAT1-2 is ricinoleate specific and enhances castor-like oil content 8

Increased hydroxy fatty acid content does not affect seed physiology 8

Co-expression castor bean PDAT1-2 and DGAT2 does not further increase the accumulation of hydroxy fatty acids in transgenic seeds 9

Discussion 9

Materials and Methods 11

Plant materials and transformation 11

Identification of three PDAT genes from castor bean 11

Cloning of castor bean PDAT genes 11

RT-PCR 12

Vector construction 12

Subcellular localization 12

Fatty acid analysis 12

TLC analysis 12

Supplementary date 12

Funding 12

Acknowledgments 13

References 13

2. Vascular-specific activity of the Arabidopsis carotenoid cleavage dioxygenase 7 gene promoter (Plant Cell Report) / Ying Shi Liang ; Yun-A Jeon ; Sun-Hyung Lim ; Jae Kwang Kim ; Jong-Yeol Lee ; Young-Mi Kim ; Yeon-Hee Lee ; Sun-Hwa Ha 16

Abstract 16

Keywords 16

Introduction 16

Materials and methods 17

RNA analysis 17

Vector construction and Arabidopsis transformation 17

Fluorometric GUS assay 18

Histochemical GUS assay 18

Results 18

Spatial expression of endogenous genes encoding four Arabidopsis CCDs 18

Organ-specific activities of the AtCcd1, 4 and 7 gene promoters 18

Constitutive and ubiquitous expression of the AtCcd7 promoter 19

Vascular specificity of the AtCcd7 promoter in trangenic Arabidopsis 19

Discussion 20

Acknowledgments 23

References 23

3. Promoters of three Brassica rapa FLOWERING LOCUS C differentially regulate gene expression during growth and development in Arabidopsis (Genes & Microbiol Biotechnol) / Joon Ki Hong ; Soo-Yun Kim ; Jung Sun Kim ; Jin A Kim ; Beom-Seok Park ; Yeon-Hee Lee 24

Abstract 24

Keywords 24

Introduction 24

Materials and Methods 25

Plant materials and growth conditions 25

Analysis of BrFLC promoters 25

Construction of promoter-GUS expression vectors 25

Plant transformation 26

Histochemical analysis 26

Semi-quantitative RT-PCR analysis 26

Results and Discussion 26

Differential activity of BrFLC family gene promoters during plant growth 26

Expression of a reporter gene induced by the BrFLC family gene promoters in vegetative tissues 27

Expression of a reporter gene induced by the BrFLC family gene promoters in reproductive organs 29

Comparison of the BrFLC promoter sequences 29

Acknowledgments 30

References 31

4. Egr-1 유전자의 발현 유도물질을 생산하는 항균성 저 영양 세균의 분리 및 동정(Isolation and Identification of Three Pseudomonas koreensis Strains with Anti-microbial Activities Producing Inducers of the Expression of Egr-l Gene (Korean J Microbiol Biotechnol) / 윤상홍 ; 김동관 ; 이영한 ; 신순영 ; 권순우 ; 이창묵 ; 강한철 ; 구본성 32

서론 32

재료 및 방법 32

사용 배지 및 균주 32

저 영양 세균 분리, 물질은행 제작, 선발 33

항균활성 검정 33

p21 및 Egr-1 luciferase 프로모터 리포터 활성 분석 33

Westem blotting 33

생화학적 분석에 의한 균 동정 34

16S rDNA 분리 및 염기서열 분석 34

결과 및 고찰 34

암 억제유전자의 발현 촉진 물질 생산 균주의 선발 34

선발 균주의 항균효과 검정 35

선발 균주의 동정 36

요약 38

Acknowledgment 38

REFERENCES 38

5. Functional characterization of a delta 6-desaturase gene from the black seabream (Acanthopagrus schlegeli) (Biotechnol Lett) / Sun Hee Kim ; Jong Bum Kim ; So Yun Kim ; Kyung Hee Roh ; Hyun Uk Kim ; Kyeong-Ryeol Lee ; Yo Soon Jang ; Mi Kwon ; Jong Sug Park 40

Abstract 40

Keywords 40

Introduction 40

Materials and methods 41

Fish meterials 41

Cloning of the full-length delta 6 desaturase cDNA from A. schlegeli 42

Vector construction and yeast transformation 42

Functional characterization of AsD6DES in Yeast 42

Fatty acid extraction and determination 42

Results 44

Isolation of AsD6DES from Acanthopagrus schlegeli 44

Characterization of the putative protein sequence of AtD6DES gene 44

Functional analysis of the A. schlegeli delta 6-fatty acid desaturase 44

Discussion 45

Acknowledgments 47

References 47

6. Expression of the Human Tissue-Plasminogen Activator in Hairy Roots of Oriental Melon (Cucumis melo) (Plant Mol Biol Rep) / Sol Kang ; Hemavathi Ajjappala ; Hyun-Hui Seo ; Joon-Soo Sim ; Sang-Hong Yoon ; Bon-Sung Koo ; Yong-Hwan Kim ; Sukchan Lee ; Bum-Soo Hahn 50

Abstract 50

Keywords 50

Introduction 50

Materials and Methods 51

Construction of Expression Vectors 51

Induction of Hairy Roots 51

PCR and RT-PCR Analysis 52

Enzyme-Linked Immunosorbent Assay 52

Western Blot Analysis 52

Fibrinolytic Activity and Inhibition of the Fibrinolytic Activity of Recombinant t-PA Using PAI-1 52

Results 52

Construction of Expression Vector and Induction of Transgenic Hairy Roots 52

Molecular Analysis of the t-PA Gene in Transgenic Hairy Roots 53

Fibrinolysis of Recombinant t-PA 53

Enzyme-Linked Immunosorbent Assay 53

ELISA 54

Western Blot Analysis 54

Inhibition of Fibrinolytic Activity of Recombinant t-PA Using PAI-1 55

Discussion 55

Acknoweledgments 56

References 56

7. Bacillus licheniformis로 부터 분리된 phospholipase D 유전자의 발현 및 생화학 특성(Expression and Biochemical Characteristics of a Phospholipase D from Bacillus licheniformis) (J Appl Bioi Chem) / 강한철 ; 윤상홍 ; 이창묵 ; 구본성 58

서론 58

재료 및 방법 59

균주 및 배양조건. 59

PLD 유전자의 클로닝. 59

PLD 유전자의 발현 및 조단백질 추출. 59

PLD의 친화 크로마토그라피 분리. 59

PLD의 효소활성 측정. 60

효소의 활성도 및 안정화도에 미치는 pH, 온도 및 양 이온의 영향. 60

기타 분석 방법. 60

결과 및 고찰 60

PLD의 E. coli 내에서의 발현 및 단백질 분리. 60

최적 pH 및 pH 안정성. 61

최적 온도 및 열 안정성. 61

PLD의 활성도에 미치는 2가 양이온 효과. 62

여러 detergent가 효소활성도에 미치는 영향. 62

초록 63

감사의 글 63

참고문헌 63

8. Inhibition of Helicobacter pylori adhesion to Kato III cells by intact and low molecular weight acharan sulfate (Glycoconj J) / Joon-Soo Sim ; Bum-Soo Hahn ; A-Rang Im ; Youmie Park ; Ji-Eun Shin ; Eun-Ah Bae ; Dong-Hyun Kim ; Yeong Shik Kim 66

Abstract 66

Keywords 66

Introduction 66

Materials and methods 67

Materials 67

Chemical depolymerization of AS 67

Agarose gel electrophoresis 67

Average MW determination of intact AS and LMWAS 68

Disaccharide compositional analysis of intact AS and LMWAS 68

¹H-NMR analysis 68

Results 70

Average MW determination of intact AS and LMWAS 70

Compositional analysis of LMWAS 71

¹H-NMR analysis 71

In vitro inhibitory effects of AS on adhesion and VacA vacuolation of H. pylori 63

Discussion 72

Acknowledgements 72

References 72

9. 식물에서 인체와 유사한 N-glycan 구조를 가진 당단백질 생산에 대한 최근 연구동향(Recent Studies on the Production of Glycoproteins Containing Humanized N-glycan Structures in Plants) (Korean J Intl Agriculture) / 심준수 ; 신동진 ; 한범수 74

식물은 74

당단백질 당화과정의 중요성 76

동물과 식물사이의 N-glycan 구조의 차이점 78

식물유래 재조합 당단백질 N-glycan 공학 78

1. 소포체로의 재조합 당단백질 표적화 78

2. 식물특이 당전이효소 유전자 발현제어 78

3. 인체특이 당전이효소 및 sialic acid 생합성 관련 유전자 삽입 79

결론 80

사사 80

인용문헌 80

10. 형질전환체에서 도입유전자 구조와 메틸화가 유전자 발현에 미치는 영향(Effects of Transgene Organization and Methylation on Gene Expression in Transgenic Plants) (Korean J Inti Agriculture) / 이연희 ; 손성한 ; 서석철 ; 윤인선 ; 김둘이 84

전세계적으로 84

재료 및 방법 85

Agrobacterium을 이용한 벼 형질전환 85

Southem과 northem 분석 85

유전자 발현 분석 85

결과 및 고찰 85

도입 유전자 구조 및 발현 분석 85

세대진전에 따른 유전자 발현 변이 분석 87

적요 89

사사 90

인용문헌 90

11. 참외모상근 내에 인체 조직형 플라스미노겐 활성화인자 변이체들의 발현(Expression of Human Tissue-Type Plasminogen Activator Derivatives in Transgenic Hairy Root of Oriental Melon) (Cucumis melo) (Korean J Intl Agriculture) / 헤마바티 ; 심준수 ; 김용환 ; 한범수 92

인체 92

재료 및 방법 93

발현 백터 제조 및 참외모상근 형질전환 93

피브린 분해활성 측정 93

ELISA 93

Westem blot 분석 94

결과 및 고찰 94

T-PA 변이체 단백질 발현 벡터 제조와 참외모상근 형질전환체 확인 94

참외 모상근 내에 t-PA 유전자 삽입 및 전사 확인 94

참외모상근 형질전환체 추출물의 피브린 분해 활성 확인 95

참외모상근 내에 존재하는 t-PA 변이체 단백질들의 발현량 측정 95

재조합 t-PA 단백질의 분자량 측정 95

적요 96

사사 96

참고문헌 96

12. 식물 지방산 생산량의 증진을 위한 생명공학 연구현황(Current biotechnology for the increase of vegetable oil yield in transgenic plants) (J Plant Biotechnol) / 이경렬 ; 최윤정 ; 김순희 ; 노경희 ; 김종범 ; 김현욱 98

Abstract 98

서론 98

TAG 구성성분의 전구체 합성 유전자의 과다발현을 통한 종자 지방산 함량 증진 100

지질합성에 관여하는 유전자의 과다발현을 통한 종자지방산 함량 증진 100

지질과 다른 대사산물간의 탄소분배 조절을 통한 종자지방산 함량 증진 102

전사인자 조절을 통한 종자지방산 함량 증진 103

전망 104

적요 105

사사 105

인용문헌 105

13. Overproduction of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) confers resistance to the herbicide glyphosate in transgenic rice (J Plant Biotechnol) / Soo In Lee ; Hyun Uk Kim ; Dongjin Shin ; Jin A Kim ; Joon Ki Hong ; Young Mi Kim ; Yeon-Hee Lee ; Bon Sung Koo ; Sun Jong Kwon ; Seok Chul Suh 108

Abstract 108

Keywords 108

Introduction 108

Material and methods 109

Construction of the CaMV35S-EPSPS gens 109

Rice transformation 109

Southern and Northern blot analysis 109

EPSPS enzyme assay 109

Herbicide application 110

Results and Discussions 110

Production of fertile transgenic rice plants 110

Inheritance of glyphosate resistance 111

Glyphosate tolerance in transgenic rice plants in the field 112

Acknowledgement 112

References 112

14. 영산 유채를 이용한 형질전환체 생산(Production of Transgenic Plants in Brassica napus Winter Cultivar 'Youngsan') (J Appl BioI Chem) / 노경희 ; 곽보경 ; 김현욱 ; 이경렬 ; 김순희 ; 서미정 ; 김효진 ; 김종범 114

서론 114

재료 및 방법 115

식물재료. 115

종자소독과 재료육성. 115

배지조제. 115

아그로박테리움 Strain과 Plasmid. 115

형질전환. 115

GUS 활성조사. 115

통계분석. 115

Southern hybridization. 116

결과 116

재분화. 116

형질전환. 116

형질전환체 육성과 유전분석. 116

고찰 118

초록 119

감사의 글 119

참고문헌 119

15. 국내 특수 생태환경의 탈질 저영양 세균의 종 다양성 및 생리적 특성 분석(Analysis of Species Variety and Physiological Characteristics of Denitrifying Oligotrophic Bacteria Isolated from the Specific Environment in Korea) (Korean J Microbiol Biotechnol) / 이창묵 ; 원항연 ; 권순우 ; 강한철 ; 구본성 ; 윤상홍 122

서론 122

재료 및 방법 123

시료 채취와 미생물 배양 배지 123

저영양 세균의 분리 123

탈질소 세균의 선발 123

탈질소 세균의 16S rRNA 유전자 염기서열 분석 124

탈질소 세균의 생존 특성 분석 124

결과 및 고찰 124

탈질소 균의 선발 124

탈질소 균의 종 다양성 분석 124

탈질소 균 속(Genus) 별 스트레스에 대한 생존 특성 분석 124

요약 128

Acknowledgement 128

REFERENCES 128

16. Identification and characterization of inhibitors of Haemophilus influenzae acetohydroxyacid synthase (Enzyme and Microbial Technology) / Vinayakumar Gedi ; Ji-Young Moon ; Won-Mook Lim ; Mi-Young Lee ; Sang-Choon Lee ; Bon-Sung Koo ; Sanjay Govindwar ; Moon-Young Yoon 130

ARTICLE INFO 130

ABSTRACT 130

1. Introduction 130

2. Materials and methods 131

2.1. Materials 131

2.2. Cloning and purification of Hi-AHAS CSU 131

2.3. Enzyme assay 131

2.4. Log P calculation and screening of Hi-AHAS inhibitors 131

2.5. Preparation of Hi-AHAS homology model 131

2.6. Molecular docking 131

3. Results and discussion 131

3.1. Expression and purification of Hi-AHAS 131

3.2. Screenign and log P calculation of Hi-AHAS inhibitors 132

3.3. Inhibition kinetics 132

3.4. Homology modeling and docking analysis 133

3.5. Binding modes of hit chemicals 133

4. Conclusion 134

Acknowledgements 134

References 134

17. 메밀을 이용한 속성장의 품질특성(Quality Characteristics of Buckwheat (Fagopyrum esculentum) Soksungjang)) (Kor J Food Sci Technol) / 최혜선 ; 이성영 ; 백성열 ; 구본성 ; 윤향식 ; 박혜영 ; 여수환 136

서론 136

재료 및 방법 137

메밀 속성장 제조 137

아미노태 질소 측정 137

총균수 측정 137

Bacillus cereus 개체수 변화 137

향기성분 137

메밀 속성장 추출물 제조 137

혈전 용해능 137

항산화도 측정 137

ACE 저해도 137

관능검사 137

결과 및 고찰 138

아미노태 질소 함량 138

총균수 측정 및 B. cereus 개체수 변화 138

향기성분 138

혈전용해능 139

항산화도 139

Angiotensin convert enzyme 저해도 139

관능검사 140

요약 140

감사의 글 140

문헌 140

18. Malonyl-CoA Synthetase, Encoded by ACYL ACTIVATING ENZYME13, Is Essential for Growth and Development of Arabidopsis (Plant Cell) / Hui Chen ; Hyun Uk Kim ; Hua Weng ; John Browse 142

INTRODUCTION 142

RESULTS 143

AAE13 Encodes a Malonyl-CoA Synthetase 143

AAE13 Is the Founding Member of a Novel Phylogenetic Clade of the Eukaryotic AAE Superfamily 143

Subcellular Localization of AAE13 145

Malonyl-CoA Synthetase Activity and Malonic Acid in Arabidopsis Tissues 145

Exogenous Malonic Acid Is Toxic to Arabidopsis 147

Compromised Growth and Development of an aae 13 Mutant 148

Complementation of aae13-1 Plants 149

Plants Overexpressing AAE13 Show Increased Sensitivity to Exogenous Malonate 150

DIUSCUSSION 151

METHODS 153

Plant Materials and Growth Conditions 153

Chemicals 153

AAE13 Phylogeny 153

Expression and Purification of Recombinant AAE13 and ACSF3 154

Measurement of Acyl-CoA Synthetase Activity 154

Metabolic Profiling of Water-Soluble Constituents in Plant Samples by GC-MS 154

Mutant Complementation 155

Subcellular Localization 155

Generation of Pro35S:AAE13 Plants 155

Accession Numbers 155

Supplemental Date 155

ACKNOWLEDGMENTS 155

REFERENCES 155

19. Novel Metagenome-Derived Carboxylesterase That Hydrolyzes β-Lactam Antibiotics (APPLIED AND ENVIRONMENTAL MICROBIOLOGY) / Jeong Ho Jeon ; Soo-Jin Kim ; Hyun Sook Lee ; Sun-Shin Cha ; Jung Hun Lee ; Sang-Hong Yoon ; Bon-Sung Koo ; Chang-Muk Lee ; Sang Ho Choi ; Sang Hee Lee ; Sung Gyun Kang ; Jung-Hyun Lee 158

MATERIALS AND METHODS 159

Strains, library construction, and screening. 159

Subcloning and sequence analysis. 159

Expression and purification of recombinant EstU1. 159

Esterase assay. 159

Biochemical properties of esterase. 159

β-Lactamase assay. 159

HPLC analysis. 159

Determination of kinetic parameters of β-lactamase activity. 159

Site-directed mutagenesis of EstU1. 160

Nucleotide sequence accession number. 160

RESULTS 160

Screening and sequence analysis of a clone with lipolytic activity. 160

Purification and characterization of EstU1. 160

Determination of β-lactamase activity. 160

Determination of kinetic parameters. 162

Site-directed mutagenesis. 162

DISCUSSION 162

ACKNOWLEDGMENTS 163

REFERENCES 163

20. Production of ketocarotenoids in transgenic carrot plants with an enhanced level of β-carotene (Plant Biotechnol Rep) / Mi-Jeong Ahn ; Seol Ah Noh ; Sun-Hwa Ha ; Kyoungwhan Back ; Shin Woo Lee ; Jung Myung Bae 166

Abstract 166

Keywords 166

Introduction 166

Materials and methods 167

Cloning of HpBkt cDNA 167

Generation of carrot transgenic plants 167

Identification of carrot transgenic plants 168

Reverse transcriptase-PCR 168

Analysis of carotenoids 168

Results 169

Cloning of HpBkt cDNA 169

Generation and identification of HpBkt-overexpressing carrot plants 169

Transcriptional regulation of endogenous carotenogenic genes in HpBkt-transgenic carrot plants 169

Phenotypic alteration in the HpBkt-transgenic carrot plants 170

Carotenoid cantent in the HpBkt-transgenic carrot plants 171

Discussion 171

Acknowledgments 172

References 172

21. Proteomic identification of rhythmic proteins in rice seedlings (Biochimica et Biophysica Acta) / Heeyoun Hwang ; Man-Ho Cho ; Bum-Soo Hahn ; Hyemin Lim ; Yong-Kook Kwon ; Tae-Ryong Hahn ; Seong Hee Bhoo 174

ARTICLE INFO 174

ABSTRACT 174

1. Introduction 174

2. Materials and methods 175

2.1. Plant materials and growth conditions 175

2.2. Protein extraction 175

2.3. 2-DE 175

2.4. MS analysis and database search 175

2.5. RT-PCR 175

3. Results 176

3.1. Diurnal rhythmic proteins 176

3.2. Rhythmic proteins under continuous dark condition 178

3.3. RT-PCR analysis of rhythmic proteins 178

3.4. Comparison of rhythmic proteome with transcriptome data 180

4. Discussion 180

Acknowledgments 182

References 182

22. Computational identification of Chinese cabbage anthocyanin-specific genes (BioChip J) / ChangKug Kim ; JinA Kim ; Shoshi Kikuchi ; JiWeon Choi ; YongKab Kim ; HyunJu Park ; YoungJoo Seol ; DongSuk Park ; JangHo Hahn ; YongHwan Kim 184

Abstract 184

Keywords: Chinese cabbage, Brassica rapa, Microarray, Anthocyanin genes, Transcription factor 184

Introduction 184

Results and Discusstion 185

Candidate genes associated with pigmentation 185

Identification of orthologs 185

Transcription factors associated with anthocyanins 185

RT-PCR analysis of selected genes 188

Conclusions 189

Materials and Methods 189

Chinese cabbage cultivars 189

Experimental design and RNA extraction 189

Microarray experiment 189

Transcription factor analysis 189

Statistical analysis 190

RT-PCR 190

Acknowledgements 191

References 191

23. Determination of lipophilic compounds in genetically modified rice using gas chromatography-time-of-flight mass spectrometry (Journal of Food Composition and Analysis) / Jae Kwang Kim ; Sun-Hwa Ha ; Soo-Yun Park ; Si Myung Lee ; Hyo Jin Kim ; Sun Hyung Lim ; Seok-Cheol Suh ; Dong Hern Kim ; Hyun Suk Cho 194

ARTICLE INFO 194

ABSTRACT 194

1. Introduction 194

2. Materials and methods 195

2.1. Rice samples 195

2.2. Reagents 195

2.3. Preparation of stock solutions and standard solutions 195

2.4. Extraction and derivatization of the samples 195

2.5. GC-FID and GC-TOFMS analysis 196

2.6. Method validation for an assay of lipid components 196

2.7. Statistical analyses 196

3. Results and discussion 196

3.1. Profile of fourteen lipophilic compounds as trimethylsilyl esters 196

3.2. Method validation for an assay of 14 lipophilic compounds in a standard mixture 197

3.3. Variation of 14 lipophilic compounds in GM rice and pigmented rice cultivars 198

4. Conclusions 200

Acknowledgments 200

References 200

24. Stable Isotope Dilution Gas Chromatography-Time-of-Flight Mass Spectrometry Method for Determining Niacin in Rice (Cereal Chem) / Jae Kwang Kim ; Soo-Yun Park ; Sun-Hwa Ha ; Si Myung Lee ; Sun Hyung Lim ; Chang Yeon Yu ; Seok-Cheol Suh ; Soon Jong Kweon 202

MATERIALS AND METHODS 202

Preparation of Stock Solutions and Standard Solutions 202

Rice Samples 202

Extraction and Derivatization of the Samples 202

Method Validation for the Nicotinic Acid Assay 203

GC-TOFMS Analysis 203

RESULTS AND DISCUSSION 203

Validation of the Overall Procedures 203

Niacin Concentrations Are Substantially Equivalent in GM and Non-GM Rice 204

CONCLUSIONS 204

ACKNOWLEDGMENTS 204

LITERATURE CITED 204

25. Construction of random sheared fosmid library from Chinese cabbage and its use for Brassica rapa genome sequencing project (Journal of Genetics and Genomics) / Tae-Ho Park ; Beom-Seok Park ; Jin-A Kim ; Joon Ki Hong ; Mina Jin ; Yong-Joo Seol ; Jeong-Hwan Mun 206

Abstract 206

1. Introduction 206

2. Materials and methods 207

2.1. Fosmid library construction 207

2.2. Fosmid clone insert sizing 207

2.3. End sequencing and in silico anchoring 208

2.4. Full sequencing of fosmid clones, sequence assembly and sequence analysis 209

3. Results 209

3.1. Fosmid library construction and characterization 209

3.2. Fosmid library screening and analysis of end sequences 209

3.3. Extension of physical contigs 210

4. Discussion 210

Acknowledgements 211

References 211

판권기 213