Identification of Helicobacter pylori HopQ as a novel T4SS-associated virulence factor:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Berlin
Mensch und Buch Verl.
2015
|
| Schriftenreihe: | Biology
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | 110 S. Ill., graph. Darst. |
| ISBN: | 9783863876289 |
Internformat
MARC
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| 005 | 20151113 | ||
| 007 | t | ||
| 008 | 151006s2015 ad|| m||| 00||| eng d | ||
| 020 | |a 9783863876289 |9 978-3-86387-628-9 | ||
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| 100 | 1 | |a Belogolova, Elena |d 1979- |e Verfasser |0 (DE-588)1077217803 |4 aut | |
| 245 | 1 | 0 | |a Identification of Helicobacter pylori HopQ as a novel T4SS-associated virulence factor |c von Elena Belogolova |
| 264 | 1 | |a Berlin |b Mensch und Buch Verl. |c 2015 | |
| 300 | |a 110 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
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| 502 | |a Zugl.: Berlin, Humboldt-Univ., Diss., 2014 | ||
| 650 | 0 | 7 | |a Helicobacter pylori |0 (DE-588)4319396-1 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Proteinsekretion |0 (DE-588)4129983-8 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Virulenzfaktor |0 (DE-588)4354782-5 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
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Datensatz im Suchindex
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|---|---|
| adam_text | TABLE OF CONTENTS
ABBREVIATIONS 6
ABSTRACT 8
ZUSAMMENFASSUNG 9
1. INTRODUCTION 11
1.1 HELICOBACTER PYLORI-, GENERAL INFORMATION 11
1.2 H. PYLORI PATHOGENESIS AND HOST RESPONSE 12
1.3 H. PYLORI INFECTION AND VIRULENCE FACTORS 15
UREASE AND FLAGELLA FACILITATE THE INITIAL STEPS OF COLONIZATION 15
ADHESINS ENABLE THE ATTACHMENT TO EPITHELIAL CELLS 16
VACUOLATION CYTOTOXLN (VACA) 17
THE CAG PATHOGENICITY ISLAND (CAGPAI) AND ITS EFFECTOR PROTEIN CAGA 18
1.4 OUTER MEMBRANE PROTEINS (OMPS) AND HOPQ: THE ROLE IN PATHOGENESIS 22
2. MATERIALS AND METHODS 24
2.1 GENERAL SUPPLIERS 24
2.2 MICROBIOLOGICAL AND CELL BIOLOGICAL METHODS 24
2.2.1 BACTERIAL STRAINS, CULTIVATION, PREPARATION OF BACTERIAL STOCKS 24
2.2.2 MAINTENANCE OF CELL CULTURES 25
2.3 STANDARD MOLECULAR BIOLOGICAL METHODS 25
2.3.1 ISOLATION OF TOTAL DNA/RNA FROM H. PYLORI 25
ISOLATION OF GENOMIC DNA FROM H. PYLORI 25
ISOLATION OF TOTAL RNA FROM H. PYLORI 26
2.3.2 PCR AND RT-PCR 26
POLYMERASE CHAIN REACTION (PCR) 26
REVERSE TRANSCRIPTION PCR (RT-PCR) 27
2.3.3 AGAROSE GEL ELECTROPHORESIS AND DNA GEL PURIFICATION 27
2.3.4 CLONING PROCEDURES 27
PLASMID ISOLATION 27
DNA RESTRICTION, LIGATION AND ETHANOL PRECIPITATION 28
DNA TRANSFORMATION INTO E. COLI AND H. PYLORI 28
2.4 STANDARD BIOCHEMICAL METHODS FOR PROTEIN ANALYSIS 29
2.4.1 PROTEIN ISOLATION 29
ISOLATION OF OUTER MEMBRANE PROTEINS (OMPS) FROM H. PYLORI 29
ISOLATION OF CHOLESTEROL-BINDING PROTEINS FROM H. PYLORI 29
TCA PROTEIN PRECIPITATION 30
2.4.2 SODIUM DODECYL SULPHATE EOLYACRYLAMIDE GEL ELECTROPHORESIS
(SDS-PAGE) 30
2.4.3 PROTEIN DETECTION 31
3
HTTP://D-NB.INFO/1077258747
COOMASSLE STAINING .. 31
IMMUNOBLOTTING (WESTERN BLOT) AND MEMBRANE STRIPPING 31
2.5 LIPID ANALYSIS: THIN LAYER CHROMATOGRAPHY (TLC) 32
2.6 SITE-DIRECTED MUTAGENESIS AND COMPLEMENTATION PROCEDURES 33
2.6.1 DELETION OF HOPQ GENE IN H. PYLORI P12 33
PREPARATION OF GENETIC CONSTRUCT FOR SITE-DIRECTED MUTAGENESIS 33
DELETION OF HOPQ IN H. PYLORI P12 35
2.6.2 COMPLEMENTATION OF THE MUTANT P12AHOPQ WITH HOPQ ALLELE I AND II
35
2.7 INFECTION ASSAYS 36
2.7.1 P65 TRANSLOCATION ASSAY 36
2.7.2 ADHERENCE ASSAY, CFU QUANTIFICATION, IMMUNOFLUORESCENCE AND PHASE-
CONTRAST MICROSCOPY 36
2.7.3 HUMMINGBIRD PHENOTYPE AND CELL SCATTERING 37
2.7.4 MOTILITY ASSAY 38
2.7.5 INTERLEUKIN 8 SECRETION 38
2.8 ELECTRON MICROSCOPY 38
2.9 IN SILICO ANALYSES OF HOPQ GENE AND ITS PRODUCT 38
2.10 IN SILICO ANALYSES OF HOPQ GENE AND HOPQ PROTEIN SEQUENCES 39
2.11 STATISTICS 39
3. RESULTS 40
3.1 IN SILICO ANALYSIS OF HOPQ GENES AND HOPQ PROTEINS FROM THE STRAINS
P12 AND
G27 40
3.2 CONSTRUCTION OF H. PYLORI HOPQ-DEFECTIVE MUTANTS 45
3.3 HOPQ-DEFICIENT MUTANT SUSTAIN ADHERENCE, MOTILITY AND MORPHOLOGY OF
THE
PARENTAL STRAIN P12 48
3.4 HOPQ DELETION IMPAIRS ACTIVATION OF NF-KB SIGNALLING PATHWAY IN THE
GASTRIC
EPITHELIAL CELLS 50
3.5 HOPQ DELETION IMPAIRS ACTIVATION OF MITOGEN-ACTIVATED PROTEIN
KINASES (MAP
KINASES) IN GASTRIC EPITHELIAL CELLS 1 54
3.6 IL-8 SECRETION UPON THE INFECTION WITH THE HOPQ DELETION MUTANT 55
3.7 IMPACT OF HOPQ DELETION ON OMP PROFILE 56
3.8 HOPQ IS ESSENTIAL FOR CAGA TRANSLOCATION 57
3.9 H. PYLORI HOPQ DELETION MUTANT CAUSES WEAK T4SS-DEPENDENT EPIGENETIC
MODULATION IN AGS CELLS 60
3.10 COMPLEMENTATION OF THE H. PYLORI P12AHOPQ MUTANT 61
3.11 THE INFLUENCE OF HOPQ ON THE T4SS ASSEMBLY 66
4
3.12 THE INFLUENCE OF HOPQ ALLELE II ON CAGA TRANSLOCATION 67
3.13 IMPACT OF HOPQ DELETION ON THE CHOLESTEROL METABOLISM OF H. PYLORI
69
4. DISCUSSION 72
5. REFERENCES 81
SUPPLEMENTARY MATERIALS 102
PUBLICATIONS 108
SELBSTSTSNDIGKEITSERKLSRUNG 109
ACKNOWLEDGEMENTS 110
5
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| author | Belogolova, Elena 1979- |
| author_GND | (DE-588)1077217803 |
| author_facet | Belogolova, Elena 1979- |
| author_role | aut |
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| dewey-full | 571.964829323 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 571 - Physiology & related subjects |
| dewey-raw | 571.964829323 |
| dewey-search | 571.964829323 |
| dewey-sort | 3571.964829323 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| format | Thesis Book |
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| indexdate | 2024-07-10T07:12:34Z |
| institution | BVB |
| isbn | 9783863876289 |
| language | English |
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| physical | 110 S. Ill., graph. Darst. |
| publishDate | 2015 |
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| series2 | Biology |
| spelling | Belogolova, Elena 1979- Verfasser (DE-588)1077217803 aut Identification of Helicobacter pylori HopQ as a novel T4SS-associated virulence factor von Elena Belogolova Berlin Mensch und Buch Verl. 2015 110 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Biology Zugl.: Berlin, Humboldt-Univ., Diss., 2014 Helicobacter pylori (DE-588)4319396-1 gnd rswk-swf Proteinsekretion (DE-588)4129983-8 gnd rswk-swf Virulenzfaktor (DE-588)4354782-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Helicobacter pylori (DE-588)4319396-1 s Proteinsekretion (DE-588)4129983-8 s Virulenzfaktor (DE-588)4354782-5 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=028336279&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Belogolova, Elena 1979- Identification of Helicobacter pylori HopQ as a novel T4SS-associated virulence factor Helicobacter pylori (DE-588)4319396-1 gnd Proteinsekretion (DE-588)4129983-8 gnd Virulenzfaktor (DE-588)4354782-5 gnd |
| subject_GND | (DE-588)4319396-1 (DE-588)4129983-8 (DE-588)4354782-5 (DE-588)4113937-9 |
| title | Identification of Helicobacter pylori HopQ as a novel T4SS-associated virulence factor |
| title_auth | Identification of Helicobacter pylori HopQ as a novel T4SS-associated virulence factor |
| title_exact_search | Identification of Helicobacter pylori HopQ as a novel T4SS-associated virulence factor |
| title_full | Identification of Helicobacter pylori HopQ as a novel T4SS-associated virulence factor von Elena Belogolova |
| title_fullStr | Identification of Helicobacter pylori HopQ as a novel T4SS-associated virulence factor von Elena Belogolova |
| title_full_unstemmed | Identification of Helicobacter pylori HopQ as a novel T4SS-associated virulence factor von Elena Belogolova |
| title_short | Identification of Helicobacter pylori HopQ as a novel T4SS-associated virulence factor |
| title_sort | identification of helicobacter pylori hopq as a novel t4ss associated virulence factor |
| topic | Helicobacter pylori (DE-588)4319396-1 gnd Proteinsekretion (DE-588)4129983-8 gnd Virulenzfaktor (DE-588)4354782-5 gnd |
| topic_facet | Helicobacter pylori Proteinsekretion Virulenzfaktor Hochschulschrift |
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