Structural insight into the Cpx-two-component system:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Göttingen
Cuvillier Verl.
2011
|
| Ausgabe: | 1. Aufl. |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | IX, 111 S. Ill., graph. Darst. |
| ISBN: | 9783869556413 |
Internformat
MARC
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| 100 | 1 | |a Zhou, Xiaohui |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Structural insight into the Cpx-two-component system |c von Xiaohui Zhou |
| 250 | |a 1. Aufl. | ||
| 264 | 1 | |a Göttingen |b Cuvillier Verl. |c 2011 | |
| 300 | |a IX, 111 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a Zugl.: Berlin, Humboldt-Univ., Diss., 2011 | ||
| 650 | 0 | 7 | |a Gram-negative Bakterien |0 (DE-588)4158020-5 |2 gnd |9 rswk-swf |
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Datensatz im Suchindex
| _version_ | 1804143825457774592 |
|---|---|
| adam_text | IMAGE 1
INDEX
ABSTRACT I
ZUSAMMENFASSUNG ILL
ABBREVIATIONS V
INDEX VII
1. INTRODUCTION 1
1.1. TWO-COMPONENT SYSTEMS 1
1.1.1. STRUCTURAL INSIGHT INTO THE FUNCTIONS OF HISTIDINE KINASES 3
1.1.2. BASIC STRUCTURES AND FUNCTIONS OF RESPONSE REGULATORS 7
1.1.3. INTERACTION BETWEEN HISTIDINE KINASE AND RESPONSE REGULATOR. 9
1.1.4. AUXILIARY PROTEINS IN TWO-COMPONENT SYSTEMS 11
1.2. THE CPX TWO COMPONENT SYSTEM 13
1.3. THE THIRD COMPONENT OF THE CPX PATHWAY - CPXP 16
1.4. THE GOALS OF MY STUDY 17
2. RESULTS 19
2.1. CRYSTALLIZATION OF THE RESPONSE REGULATOR CPXR 19
2.2. CPXP STRUCTURE AND ITS FUNCTIONS IN CPX PATHWAY 23
2.2.1. PURIFICATION AND CRYSTALLIZATION OF CPXP 23
2.2.2. SECONDARY STRUCTURE PREDICTION OF CPXP 26
2.2.3. COMPARISON OF WILD-TYPE CPXP AND CPXPAL51 28
2.2.4. CRYSTALLIZATION OF CPXPAL 51 31
2.2.5. PURIFICATION AND OPTIMIZATION OF SE-MET CPXPA151 CRYSTALS 33
2.2.6. X-RAY DATASETS COLLECTION OF CPXPAL 51 AND SE-MET CPXPAL 51
CRYSTALS 35
2.2.7. OVERALL STRUCTURE OF CPXPA151 39
2.2.8. DIMER STRUCTURE OF CPXPAL 51 AND ITS STABILITY 40
2.2.9. CPXP SUSCEPTIBILITY FOR DEGRADATION IS DEPENDENT ON ITS
C-TERMINUS
45
2.2.10. INTERACTION OF CPXP WITH CPXA 47
2.2.11. IMPACT OF THE C-TERMINUS OF CPXP ON ITS FUNCTION 52
2.2.12. CHAPERONE-LIKE STRUCTURE OF CPXP 54
2.2.13. CRYSTALS OF CPXPAL 51 VARIANTS E79R/R144E AND R144E 58
VII
BIBLIOGRAFISCHE INFORMATIONEN HTTP://D-NB.INFO/1010010611
DIGITALISIERT DURCH
IMAGE 2
INDEX
3. DISCUSSION 61
3.1. THE STRUCTURE OF CPXP 62
3.2. INHIBITORY FUNCTION OF CPXP 63
3.3. QUALITY CONTROL FUNCTION OF CPXP 65
3.4. A MODEL FOR SIGNAL INTEGRATION BY CPXP 67
3.5. THE CPX ENVELOPE STRESS PATHWAY: A POTENTIAL DRUG TARGET IN
PATHOGENIC BACTERIA 68
4. MATERIALSAND METHODS 71
4.1. BACTERIAL STRAINS, PLASMIDS AND PRIMERS 71
4.2. MEDIA AND PROTOCOLS FOR CULTIVATION OF COLI 77
4.3. WORKING WITH DNA 78
4.3.1. ISOLATION OF PLASMID DNA FROM E. COLI 78
4.3.2. ISOLATION OF GENOMIC DNA FROM E. COLI 79
4.3.3. GEL ELECTROPHORESIS OF DNA 79
4.3.4. EXTRACTION OF DNA FROM AGAROSE GELS 79
4.3.5. SITE-DIRECTED MUTAGENESIS 79
4.3.6. AMPLIFICATION OF DNA 80
4.3.7. RESTRICTION, LIGATION AND SEQUENCING OF DNA 80
4.3.8. PREPARATION OF COMPETENT CELLS AND TRANSFORMATION 81
4.4. PROTEIN PURIFICATION AND PRODUCTION 83
4.4.1. EXPRESSION OF PROTEINS AND PREPARATION OF CYTOSOLIC FRACTION 83
4.4.2. PURIFICATION OF RESPONDS REGULATOR CPXR 83
4.4.3. PURIFICATION OF HISTIDINE KINASE CPXA SENSOR DOMAIN 84
4.4.4. PURIFICATION OF CPXP, CPXPA151 AND ITS VARIANTS 85
4.5. PROTEIN ANALYSIS MANIPULATION 86
4.5.1. ANALYSIS OF PROTEIN CONCENTRATION 86
4.5.2. SDS-PAGE AND COOMASSIE STAINING 86
4.5.3. LIMITED PROTEOLYSIS OF PROTEINS 88
4.5.4. CD SPECTRA 88
4.5.5. WESTERN BLOTTING 89
4.5.6. SS-GALACTOSIDASE ACTIVITY ANALYSIS 90
VIII
IMAGE 3
INDEX
4.6. BIOCHEMISTRY METHODS 91
4.6.1. CELL FRACTIONATIONS AND DETERMINATION OF CPXP EXPRESSION 91
4.6.2. SIZE EXCLUSION CHROMATOGRAPHY (SEC) 91
4.6.3. PROTEIN CRYSTALLIZATION 92
4.6.4. DATA COLLECTION AND STRUCTURE DETERMINATION 93
4.7. ANTIBODY PREPARATION OF CPXP AND CPXR 94
5. REFERENCES 95
ACKNOWLEDGEMENT 111
IX
|
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| author_facet | Zhou, Xiaohui |
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| dewey-tens | 570 - Biology |
| discipline | Biologie |
| edition | 1. Aufl. |
| format | Thesis Book |
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| isbn | 9783869556413 |
| language | English |
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| physical | IX, 111 S. Ill., graph. Darst. |
| publishDate | 2011 |
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| spelling | Zhou, Xiaohui Verfasser aut Structural insight into the Cpx-two-component system von Xiaohui Zhou 1. Aufl. Göttingen Cuvillier Verl. 2011 IX, 111 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zugl.: Berlin, Humboldt-Univ., Diss., 2011 Gram-negative Bakterien (DE-588)4158020-5 gnd rswk-swf Zweikomponentensystem Molekularbiologie (DE-588)7710598-9 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Gram-negative Bakterien (DE-588)4158020-5 s Zweikomponentensystem Molekularbiologie (DE-588)7710598-9 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=021137277&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Zhou, Xiaohui Structural insight into the Cpx-two-component system Gram-negative Bakterien (DE-588)4158020-5 gnd Zweikomponentensystem Molekularbiologie (DE-588)7710598-9 gnd |
| subject_GND | (DE-588)4158020-5 (DE-588)7710598-9 (DE-588)4113937-9 |
| title | Structural insight into the Cpx-two-component system |
| title_auth | Structural insight into the Cpx-two-component system |
| title_exact_search | Structural insight into the Cpx-two-component system |
| title_full | Structural insight into the Cpx-two-component system von Xiaohui Zhou |
| title_fullStr | Structural insight into the Cpx-two-component system von Xiaohui Zhou |
| title_full_unstemmed | Structural insight into the Cpx-two-component system von Xiaohui Zhou |
| title_short | Structural insight into the Cpx-two-component system |
| title_sort | structural insight into the cpx two component system |
| topic | Gram-negative Bakterien (DE-588)4158020-5 gnd Zweikomponentensystem Molekularbiologie (DE-588)7710598-9 gnd |
| topic_facet | Gram-negative Bakterien Zweikomponentensystem Molekularbiologie Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=021137277&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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