The 6C RNA of Corynebacterium glutamicum:
Saved in:
| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
Jülich
Forschungszentrum Jülich, Zentralbibliothek
2014
|
| Series: | Schriften des Forschungszentrums Jülich
Reihe Gesundheit ; 76 |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | 144 S. Ill., graph. Darst. |
| ISBN: | 9783958060036 |
Staff View
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Record in the Search Index
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| adam_text | CONTENT
SUMMARY 1
ZUSAMMENFASSUNG 3
1 INTRODUCTION 5
1.1 CORYNEBACTERIUM GLUTAMICUM, A MODEL ORGANISM OF BIOTECHNOLOGICAL
RELEVANCE 5
1.2 SMALL RNAS IN BACTERIA 7
1.2.1 C/S-ENCODED ANTISENSE RNAS 8
1.2.2 TRANS-ENCODED ANTISENSE RNAS 10
1.2.3 SRNAS THAT MODULATE PROTEIN ACTIVITY 12
1.3 THE 6C RNA FAMILY 14
1.4 AIMS OF THIS WORK 17
2 MATERIALS AND METHODS 19
2.1 BUFFERS AND ANTIBIOTICS 19
2.2 CULTURE MEDIA 21
2.3 OLIGONUCLEOTIDES 22
2.4 BACTERIAL STRAINS AND PLASMIDS 24
2.5 PLASMID CONSTRUCTIONS 27
2.5.1 CONSTRUCTION OF PK19MOBSACS DERIVATIVES FOR THE DELETION OF GENES
27
2.5.2 CONSTRUCTION OF PAN AND PJC1 DERIVATIVES AS EXPRESSION PLASMIDS
FOR C. GLUTAMICUM 28
2.5.3 CONSTRUCTION OF PUCBM21 DERIVATIVES AS EXPRESSION PLASMIDS FOR E.
COLI 28
2.6 CULTIVATION OF BACTERIA 28
2.6.1 CULTIVATION OF E. COLI 28
2.6.2 CULTIVATION OF C. GLUTAMICUM 29
2.6.3 MAINTENANCE OF BACTERIA 29
2.6.4 DETERMINATION OF BACTERIAL GROWTH 30
2.6.5 DETERMINATION OF THE CELL DRY WEIGHT (CDW) 30
2.6.6 CULTIVATION IN MICROTITER SCALE IN THE BIOLECTOR SYSTEM 30
2.7 MOLECULAR BIOLOGY METHODS 31
2.7.1 ISOLATION OF NUCLEIC ACID 31
2.7.2 NUCLEIC ACID GEL ELECTROPHORESIS 32
2.7.3 DETERMINATION OF NUCLEIC ACID CONCENTRATIONS 33
2.7.4 POLYMERASE CHAIN REACTION (PCR) 33
2.7.5 PURIFICATION OF DNA FRAGMENTS 34
2.7.6 RECOMBINANT DNA TECHNIQUES 34
2.7.7 DNA SEQUENCING 35
HTTP://D-NB.INFO/1065757050
2.7.8 GENERATION AND TRANSFORMATION OF COMPETENT . COLI 35
2.7.9 GENERATION AND TRANSFORMATION OF COMPETENT C. GLUTAMICUM 36
2.7.10 CHROMOSOMAL GENE REPLACEMENT USING THE PK19MO SACB SYSTEM 36
2.7.11 RNA IN VITRO TRANSCRIPTION 37
2.7.12 RNA PURIFICATION BY PHENOL CHLOROFORM EXTRACTION 37
2.7.13 ETHANOL PRECIPITATION OF RNA 38
2.7.14 NORTHERN BLOT ANALYSIS 38
2.7.15 QUANTIFICATION OF 6C RNA MOLECULES PER CELL 39
2.7.16 RNA-PROTEIN INTERACTION STUDIES 39
2.8 GLOBAL GENE EXPRESSION ANALYSIS USING DNA MICROARRAYS 41
2.8.1 SYNTHESIS AND LABELING OF CDNA 41
2.8.2 C. GLUTAMICUM DNA MICROARRAY HYBRIDIZATION 42
2.8.3 DNA MICROARRAY FLUORESCENCE SIGNAL MEASUREMENT AND DATA ANALYSIS
43
2.9 PROTEIN BIOCHEMICAL METHODS 44
2.9.1 CELL DISRUPTION METHODS 44
2.9.2 DETERMINATION OF PROTEIN CONCENTRATIONS 44
2.9.3 PROTEIN PRECIPITATION 45
2.9.4 SDS POLYACRYLAMIDE GEL ELECTROPHORESIS ! 45
2.9.5 PROTEIN OVEREXPRESSION AND PURIFICATION 45
2.9.6 CONCENTRATION AND DESALTING OF PROTEINS 47
2.9.7 DNA-PROTEIN INTERACTION STUDIES 47
2.9.8 PGKCG ENZYME ACTIVITY ASSAYS 48
2.10 MALDI TOF MASS SPECTROMETRY 49
2.11 MICROSCOPY TECHNIQUES 50
2.12 QUANTIFICATION OF METABOLITES 50
2.12.1 QUANTIFICATION OF GLUCOSE AND ORGANIC ACIDS IN CULTURE
SUPERNATANTS 50
2.12.2 QUANTIFICATION OF AMINO ACIDS 51
3 RESULTS 53
3.1 THE 6C RNA AND ITS EXPRESSION IN C. GLUTAMICUM 53
3.1.1 THE 6C RNA IS VERY STABLE 54
3.1.2 NUMBER OF 6C RNA MOLECULES PER CELL 55
3.1.3 PUTATIVE TRANSCRIPTIONAL REGULATORS OF THE 6C RNA 56
3.2 CONSTRUCTION AND CHARACTERIZATION OF 6C RNA DELETION MUTANTS 60
3.2.1 TRANSCRIPTOME ANALYSIS OF THE A6C MUTANT 62
3.2.2 SCREENING FOR A PHENOTYPE OF THE A6C RNA MUTANT 67
3.3 PROPERTIES OF THE A6CIP MUTANT IN THE PRESENCE OF MITOMYCIN C (MMC)
69
3.3.1 GROWTH AND CELL SHAPE OF A6CIP MUTANT CULTIVATED WITH MMC 69
3.3.2 TRANSCRIPTOME ANALYSIS OF THE A6CIP MUTANT CULTIVATED WITH MMC 71
3.3.3 THE SOS RESPONSE IS ALTERED IN THE A6CIP MUTANT 74
3.3.4 COMPLEMENTATION OF THE A6CIP MUTANT MMC-PHENOTYPE 76
3.4 OVEREXPRESSION OF THE 6C RNA IN C. GLUTAMICUM PJC1-6C (+/-MMC) 78
3.4.1 GROWTH AND CELL SHAPE OF C. GLUTAMICUM PJC1-6C (+/-MMC) 78
3.4.2 TRANSCRIPTOME ANALYSIS OF C. GLUTAMICUM PJC1-6C (+/- MMC) 80
3.5 PUTATIVE MRNA TARGETS OF THE 6C RNA 87
3.5.1 DELETION AND OVEREXPRESSION OF WHCD 87
3.5.2 CHARACTERIZATION OF THE DOWNSTREAM GENES OF THE 6C RNA 91
3.6 PUTATIVE PROTEIN INTERACTION PARTNERS OF THE 6C RNA 92
3.7 EFFECT OF 6C RNA ON AMINO ACID PRODUCTION IN C. GLUTAMICUM 95
4 DISCUSSION 99
4.1 THE 6C RNA IS AN ABUNDANT AND VERY STABLE SRNA IN C. GLUTAMICUM 99
4.2 THE POTENTIAL ROLE OF THE 6C RNA UNDER SOS RESPONSE-INDUCING
CONDITIONS 100
4.3 POTENTIAL MRNA TARGETS OF THE 6C RNA 105
4.3.1 THE DOWNSTREAM GENES OF THE 6C RNA (CG0362 TO CG0369) 108
4.3.2 THE WHCD REGULATOR OF C. GLUTAMICUM 109
4.4 3-PHOSPHOGLYCERATE KINASE - A CANDIDATE PROTEIN INTERACTION PARTNER
OF THE 6C RNA111
5 REFERENCES 115
6 APPENDIX 127
6.1 NUMBER OF 6C RNA MOLECULES PER CELL 127
6.2 PUTATIVE TRANSCRIPTIONAL REGULATORS OF THE 6C RNA 128
6.3 THE SOS RESPONSE IS ALTERED IN THE A6CIP MUTANT 130
6.4 CHARACTERIZATION OF THE DOWNSTREAM GENES OF THE 6C RNA 132
6.5 DELETION AND OVEREXPRESSION OF WHCD 132
6.6 EFFECT OF 6C RNA ON AMINO ACID PRODUCTION IN C. GLUTAMICUM 134
6.7 TRANSCRIPTOME ANALYSIS 135
|
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| author_GND | (DE-588)1064188621 |
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| bvnumber | BV042427033 |
| ctrlnum | (OCoLC)905347213 (DE-599)DNB1065757050 |
| dewey-full | 579.373 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 579 - Microorganisms, fungi & algae |
| dewey-raw | 579.373 |
| dewey-search | 579.373 |
| dewey-sort | 3579.373 |
| dewey-tens | 570 - Biology |
| discipline | Biologie Geographie |
| format | Thesis Book |
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| indexdate | 2024-07-10T01:21:19Z |
| institution | BVB |
| isbn | 9783958060036 |
| language | English |
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| physical | 144 S. Ill., graph. Darst. |
| publishDate | 2014 |
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| series | Schriften des Forschungszentrums Jülich |
| series2 | Schriften des Forschungszentrums Jülich : Reihe Gesundheit |
| spelling | Pahlke, Jennifer Verfasser (DE-588)1064188621 aut The 6C RNA of Corynebacterium glutamicum Jennifer Pahlke Jülich Forschungszentrum Jülich, Zentralbibliothek 2014 144 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Schriften des Forschungszentrums Jülich : Reihe Gesundheit 76 Zugl.: Düsseldorf, Univ., Diss., 2014 (DE-588)4113937-9 Hochschulschrift gnd-content Schriften des Forschungszentrums Jülich Reihe Gesundheit ; 76 (DE-604)BV035800060 76 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027862396&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Pahlke, Jennifer The 6C RNA of Corynebacterium glutamicum Schriften des Forschungszentrums Jülich |
| subject_GND | (DE-588)4113937-9 |
| title | The 6C RNA of Corynebacterium glutamicum |
| title_auth | The 6C RNA of Corynebacterium glutamicum |
| title_exact_search | The 6C RNA of Corynebacterium glutamicum |
| title_full | The 6C RNA of Corynebacterium glutamicum Jennifer Pahlke |
| title_fullStr | The 6C RNA of Corynebacterium glutamicum Jennifer Pahlke |
| title_full_unstemmed | The 6C RNA of Corynebacterium glutamicum Jennifer Pahlke |
| title_short | The 6C RNA of Corynebacterium glutamicum |
| title_sort | the 6c rna of corynebacterium glutamicum |
| topic_facet | Hochschulschrift |
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