Studies on the chemotaxis network in Halobacterium salinarum and Helicobacter pylori:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
2002
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | München, Univ., Fak. für Chemie u. Pharmazie, Diss., 2003 |
| Beschreibung: | 174 Bl. graph. Darst. |
Internformat
MARC
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| 100 | 1 | |a Schaer, Andreas |d 1971- |e Verfasser |0 (DE-588)124498809 |4 aut | |
| 245 | 1 | 0 | |a Studies on the chemotaxis network in Halobacterium salinarum and Helicobacter pylori |c von Andreas Schaer |
| 264 | 1 | |c 2002 | |
| 300 | |a 174 Bl. |b graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a München, Univ., Fak. für Chemie u. Pharmazie, Diss., 2003 | ||
| 650 | 0 | 7 | |a Helicobacter pylori |0 (DE-588)4319396-1 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Halobacterium salinarium |0 (DE-588)4158862-9 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Chemotaxis |0 (DE-588)4127969-4 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Halobacterium salinarium |0 (DE-588)4158862-9 |D s |
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Datensatz im Suchindex
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| adam_text |
1. CONTENTS
1
1. CONTENTS
1. CONTENTS.1
2. ABBREVIATIONS.5
3. INTRODUCTION.7
3.1. THE
E. COLI CHEMOTAXIS NETWORK.7
3.2. NOVEL CHEMOTAXIS PROTEINS OUTSIDE THE
ENTEROBACTERIACEAE.10
3.3. HALOBACTERIUM SALINARUM AND
HELICOBACTER PYLORI CHEMOTAXIS.13
3.3.1. CHEMOTAXIS NETWORK IN
HALOBACTERIUM SALINARUM.13
3.3.2.
HELICOBACTER PYLORI - CHEMOTAXIS OF AN IMPORTANT HUMAN
PATHOGEN. 16
4. AIMS OF THIS STUDY.19
5. RESULTS.21
5.1. IDENTIFICATION AND EXPRESSION OF SOLUBLE MCP HOMOLOGS FROM
H.
SALINARUM
.21
5.2. PURIFICATION OF CAR AND HTRL5. 24
5.3. CHARACTERIZATION OF CAR BY CD SPECTROSCOPY.26
5.4. HEAT DENATURATION OF CAR.32
5.5. CHARACTERIZATION OF CAR BY
1H-NMR.33
5.6. CRYSTALLIZATION TRIALS FOR CAR.35
5.7. EXPRESSION, PURIFICATION AND CRYSTALLIZATION TRIALS WITH CAR
FRAGMENTS. .36
5.8. EXPRESSION OF
H. SALINARUM CHEB AND CHER IN E. COLI.37
5.9. SOLUBLE RECEPTOR HOMOLOGS IN C.
JEJUNI AND H. PYLORI.38
5.10. CRYSTALLIZATION OF CHEMOTAXIS COMPONENTS HP0599, CJ0448 AND
CHEW.42
5.11. THE CHEMOTAXIS NETWORK IN
H. PYLORI.46
5.11.1. MODULATION OF CHEF AUTOPHOSPHOYLATION ACTIVITY.46
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/967172179
1. CONTENTS
2
5.11.2. PHOSPHOTRANSFER REACTIONS FROM CHEFTO RESPONSE REGULATORS.57
5.11.3. PHOSPHATE GROUP TRANSFER THROUGH THE CHE PROTEIN NETWORK.58
5.11.4. AUTOPHOSPHORYLATION OF CHEF:D729K MUTANT.60
5.11.5. PHOSPHOTRANSFER REACTIONS FROM CHEF:D729K-P
( TO RESPONSE
REGULATORS.62
5.11.6. DECAY OF CHEY-PHOSPHATE.67
5.11.7. REVERSE PHOSPHOTRANSFER FROM CHEY TO THE KINASE.69
5.11.8. PHOSPHORYLATION OF RESPONSE REGULATORS BY ACETYL PHOSPHATE
.72
5.12. EXPRESSION AND PURIFICATION OF THE MOTOR SWITCH PROTEIN FLIM.74
5.13. DETECTION OF
H. PYLORI CHEMOTAXIS PROTEINS BY ANTISERA.79
6. DISCUSSION. 83
6.0. SOLUBLE MCP HOMOLOGUES IN TWO-COMPONENT CASCADES .83
6.1. RECOMBINANT PRODUCTION OF HALOPHILIC SIGNALING COMPONENTS
IN E. COLI.84
6.2. THE ARGININE RECEPTOR CAR - A MOLTEN GLOBULE LIKE RECEPTOR?.85
6.3. ROLE OF EUBACTERIAL SOLUBLE RECEPTORS.87
6.4. POSSIBLE ROLE OF CHEV PROTEINS. 94
6.5. THE
H. PYLORI MOTOR AND ITS SWITCH .99
6.6. ADAPTATION IN BIOLOGICAL NETWORKS AND CHEMOTAXIS IN
H. PYLORI.103
6.7. GENE REGULATION AND ADAPTATION IN A STABLE ENVIRONMENT.105
7. MATERIALS AND METHODS. 109
7.1. BACTERIAL STRAINS AND PLASMIDS.109
7.2. GENERAL MOLECULAR BIOLOGICAL TECHNIQUES.110
7.2.1. CULTURE OF
E. COLI XL1 BLUE CELLS FOR PLASMID GROWTH.110
7.2.2. PURIFICATION OF PLASMID DNA FROM
E. COLI CELLS.110
7.2.3. GENERAL PCR PROTOCOL FOR DNA AMPLIFICATION.ILL
7.2.4. PURIFICATION OF DNA AFTER ENZYMATIC REACTIONS.ILL
7.2.5. PREPARATION OF VECTOR DNA FOR LIGATION REACTIONS.112
7.2.6. PREPARATION OF PCR-DERIVED DNA FRAGMENTS FOR LIGATION
REACTIONS. 112
1. CONTENTS
3
7.2.7. LIGATION OF DNA MOLECULES.113
7.2.8. RESTRICTION ANALYSIS OF PLASMID DNA.113
7.2.9. ANALYSIS OF DNA BY AGAROSE GEL ELECTROPHORESIS.113
7.2.10. SEQUENCING OF PLASMID DNA.114
7.2.11. PREPARATION OF SHORT SYNTHETIC DSDNA LINKER.115
7.2.12. CONSTRUCTION OF MUTANT GENES BY SITE-DIRECTED MUTAGENESIS. 115
7.2.13. CONSTRUCTION OF MUTANT GENES BY PCR.116
7.2.14. CONSTRUCTION OF CHIMERIC FLIM.116
7.2.15. SYNTHESIS OF
H. SALINARUM CHER AND CHEB GENES.117
7.2.16. CONSTRUCTION OF N- AND C-TERMINAL FRAGMENTS OF CAR.119
7.2.17. PREPARATION OF ELECTROCOMPETENT
E. COLI CELLS .120
7.2.18. TRANSFORMATION OF
E. COLI CELLS.R.121
7.2.19. PRIMER DESIGN FOR PCR AND SEQUENCING REACTIONS.121
7.3. PROTEIN CHEMICAL METHODS.122
7.3.1. TEST FOR PET28A(+) DIRECTED GENE EXPRESSION.122
7.3.2. ISOLATION OF INCLUSION BODIES FROM
E. COLI CELLS.123
7.3.3. HETEROLOGOUS EXPRESSION OF PROTEINS IN
E. COLI. 123
7.3.4. PURIFICATION OF OVEREXPRESSED PROTEINS FROM
E. COLI.124
7.3.5. SDS-PAGE ANALYSIS OF PROTEINS.128
7.3.6. DETECTION OF PROTEINS AFTER SDS-PAGE BY COOMASSIE
BRILLIANT BLUE.;.129
7.3.7. DETECTION OF PROTEINS AFTER SDS-PAGE BY SILVER STAINING.129
7.3.8. WESTERN TRANSFER OF PROTEINS SEPARATED BY SDS-PAGE TO PVDF
MEMBRANE. 130
7.3.9. IMMUNODETECTION OF PROTEINS IMMOBILIZED ON PVDF MEMBANES 131
7.3.10. PRODUCTION OF ANTISERA AGAINST
H. PYLORI CHEMOTAXIS PROTEINS IN
RABBITS.132
7.3.11. PROTEIN CONCENTRATION ASSAY.133
7.4. AUTOPHOSPHORYLATION AND PHOSPHOTRANSFER REACTIONS.133
7.4.1. SYNTHESIS OF [
32P] LABELED DILITHIUM ACETYL PHOSPHATE.133
1. CONTENTS
4
7.4.2. ACYL PHOSPHATE ASSAY.134
7.4.3. PREPARATION OF PHOSPHORYLATED CHEF:D729K. 134
7.4.4. PHOSPHOTRANSFER ASSAY FROM PHOSPHORYLATED CHEF:D729K TO
RESPONSE REGULATORS.135
7.4.5. PHOSPHOTRANSFER ASSAY FROM PHOSPHORYLATED RESPONSE REGULATOR
CHEY:K106R TO CHEF:D729K.136
7.4.6. CHEF AUTOPHOSPHORYLATION AND TRANSPHOSPHORYLATION ACTIVITIES
USING [Y
32P]-ATP.136
7.4.7. RESPONSE REGULATOR AUTOPHOSPHORYLATION BY ACETYL PHOSPHATE. 137
7.4.8. QUANTITATION OF RADIOLABEL ON CHEF AFTER SDS-PAGE BY LIQUID
SCINTILLATION COUNTING.137
7.4.9. QUANTITATION OF PHOSPHOTRANSFER REACTIONS BY "PHOSPHOIMAGING"
.138
7.5. ATPASE ASSAY. 138
7.6. CIRCULAR DICHROIC MEASUREMENTS.136
7.7.
1H-NMR SPECTROSCOPY WITH CAR.139
7.8. PROTEIN CRYSTALLOGRAPHY.140
8. REFERENCES.143
APPENDIX A: EQUIPMENT. 163
APPENDIX B: PRIMERS .167
APPENDIX C: GENE SEQUENCES OF SYNTHETIC GENES.171
9. SUMMARY.173 |
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| spelling | Schaer, Andreas 1971- Verfasser (DE-588)124498809 aut Studies on the chemotaxis network in Halobacterium salinarum and Helicobacter pylori von Andreas Schaer 2002 174 Bl. graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Univ., Fak. für Chemie u. Pharmazie, Diss., 2003 Helicobacter pylori (DE-588)4319396-1 gnd rswk-swf Halobacterium salinarium (DE-588)4158862-9 gnd rswk-swf Chemotaxis (DE-588)4127969-4 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Halobacterium salinarium (DE-588)4158862-9 s Chemotaxis (DE-588)4127969-4 s DE-604 Helicobacter pylori (DE-588)4319396-1 s DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010286242&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schaer, Andreas 1971- Studies on the chemotaxis network in Halobacterium salinarum and Helicobacter pylori Helicobacter pylori (DE-588)4319396-1 gnd Halobacterium salinarium (DE-588)4158862-9 gnd Chemotaxis (DE-588)4127969-4 gnd |
| subject_GND | (DE-588)4319396-1 (DE-588)4158862-9 (DE-588)4127969-4 (DE-588)4113937-9 |
| title | Studies on the chemotaxis network in Halobacterium salinarum and Helicobacter pylori |
| title_auth | Studies on the chemotaxis network in Halobacterium salinarum and Helicobacter pylori |
| title_exact_search | Studies on the chemotaxis network in Halobacterium salinarum and Helicobacter pylori |
| title_full | Studies on the chemotaxis network in Halobacterium salinarum and Helicobacter pylori von Andreas Schaer |
| title_fullStr | Studies on the chemotaxis network in Halobacterium salinarum and Helicobacter pylori von Andreas Schaer |
| title_full_unstemmed | Studies on the chemotaxis network in Halobacterium salinarum and Helicobacter pylori von Andreas Schaer |
| title_short | Studies on the chemotaxis network in Halobacterium salinarum and Helicobacter pylori |
| title_sort | studies on the chemotaxis network in halobacterium salinarum and helicobacter pylori |
| topic | Helicobacter pylori (DE-588)4319396-1 gnd Halobacterium salinarium (DE-588)4158862-9 gnd Chemotaxis (DE-588)4127969-4 gnd |
| topic_facet | Helicobacter pylori Halobacterium salinarium Chemotaxis Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010286242&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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