The role of the secreted glycoprotein G (gG) in equine herpesvirus type 1 (EHV-1) immune modulation and virulence:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Berlin
Mensch und Buch Verl.
2011 [erschienen] 2012
|
| Schlagworte: | |
| Online-Zugang: | kostenfrei Inhaltsverzeichnis |
| Beschreibung: | Nebentitel: Immunmodulation und Beeinflussung der Virulenz durch das Glykoprotein G (gG) des Equinen Herpesvirus vom Typ 1 |
| Beschreibung: | VIII, 105, II S. Ill., graph. Darst. |
| ISBN: | 9783863870980 |
Internformat
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| 245 | 1 | 0 | |a The role of the secreted glycoprotein G (gG) in equine herpesvirus type 1 (EHV-1) immune modulation and virulence |c vorgelegt von Nora Mariella Thormann |
| 246 | 1 | 3 | |a Immunmodulation und Beeinflussung der Virulenz durch das Glykoprotein G (gG) des Equinen Herpesvirus vom Typ 1 |
| 264 | 1 | |a Berlin |b Mensch und Buch Verl. |c 2011 [erschienen] 2012 | |
| 300 | |a VIII, 105, II S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
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| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Nebentitel: Immunmodulation und Beeinflussung der Virulenz durch das Glykoprotein G (gG) des Equinen Herpesvirus vom Typ 1 | ||
| 502 | |a Berlin, Freie Univ., Diss., 2011 | ||
| 650 | 7 | |a Equine herpesvirus 1 |2 cabt | |
| 650 | 7 | |a Equine herpesvirus 4 |2 cabt | |
| 650 | 7 | |a horse diseases |2 cabt | |
| 650 | 7 | |a immune evasion |2 cabt | |
| 650 | 7 | |a glycoproteins |2 cabt | |
| 650 | 7 | |a neutrophils |2 cabt | |
| 650 | 7 | |a chemotaxis |2 cabt | |
| 650 | 7 | |a recombinatio |2 cabt | |
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| 650 | 0 | 7 | |a Pferdekrankheit |0 (DE-588)4045515-4 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Virulenz |0 (DE-588)4129834-2 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Glykoproteine |0 (DE-588)4071905-4 |2 gnd |9 rswk-swf |
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Datensatz im Suchindex
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| adam_text | IMAGE 1
TABLE OF CONTENTS
TABLE OF CONTENTS
TABLE OF CONTENTS I
LIST OF ABBREVIATIONS V
LIST OF FIGURES AND TABLES VI
ZUSAMMENFASSUNG 1
SUMMARY 3
1. INTRODUCTION 4
1.1. ANIMAL HERPESVIRUSES 4
1.1.1. VIRION ARCHITECTURE 4
1.2. EQUINE HERPESVIRUSES TYPE 1 AND 4 5
1.2.1. CLASSIFICATION AND STRUCTURE OF EQUINE HERPESVIRUS TYPE 1 AND 4 5
1.2.2. GLYCOPROTEINS IN THE VIRAL ENVELOPE 7
1.2.3. THE GLYCOPROTEIN G 7
A. BHV-1 GG AND BHV-5 GG 8
B. EHV-3 GG 8
C. FEHV-1 GG (VCKBP) 8
D. I LTV GG (VCKBP) 8
E. PRVGG (VCKBP) 9
F. EHV-1 GG (VCKBP) AND EHV-4 GG 9
1.2.4. PATHOGENESIS OF EHV-1 AND EHV-4 INFECTION 12
1.2.5. DIAGNOSIS OF EHV-1 AND EHV-4 INFECTION 12
1.2.6. CURRENT VACCINES FOR EHV-1 AND EHV-4 13
1.3. BACTERIAL ARTIFICIAL CHROMOSOMES (BACS) 14
1.3.1. BACS IN GENERAL 14
1.3.2. BACS IN VIROLOGY 14
1.4. CHEMOKINES 15
1.5. VIRAL CHEMOKINE BINDING PROTEINS OF OTHER VIRUSES 15
1.6. MURINE RESPIRATORY MODEL 17
1.7. HYPOTHESIS 18
2. MATERIALS AND METHODS 19
2.1. MATERIALS 19
2.1.1. BUFFERS AND SOLUTIONS 19
2.1.2. ANTIBIOTICS 22
2.1.3. MEDIA AND SUPPLEMENTS 22
2.1.4. CHEMICALS AND BIOCHEMICALS 23
2.1.5. ENZYMES 24
I
HTTP://D-NB.INFO/1020073020
IMAGE 2
TABLE OF CONTENTS
2.1.6. EQUIPMENT 24
2.1.7. CONSUMABLES 27
2.1.8. KITS 28
2.1.9. DNA AND PROTEIN MARKER 28
2.1.10. STAINING 29
2.1.11. CELLS 33
2.1.12. VIRUSES AND BACTERIAL ARTIFICIAL CHROMOSOMES (BACS) 33
2.1.13. PLASMIDS 34
2.1.14. ANTIBODIES 3 4
2.1.14.1. WESTERN BLOT 35
2.1.14.2. INDIRECT IMMUNOFLUORESCENCE ASSAYS (IFA) 35
2.1.14.3. FLOW CYTOMETRY 35
2.1.15. CHEMOKINES 36
2.1.16. ANIMALS 36
2.2. METHODS 3 7
2.2.1. CELL CULTURE 37
A. MAINTENANCE OF CELLS 37
B. CELL COUNTING 37
C. CRYOCONSERVATION 37
D. TRANSFECTION AND CONTRANSFECTION 37
2.2.2. VIROLOGICAL METHODS 38
A. PREPARATION OF VIRAL STOCKS 38
B. TITRATION 38
C. SINGLE-STEP GROWTH KINETICS 39
D. IMMUNOFLUORESCENCE ASSAY (IFA) 39
2.2.3. MOLECULAR METHODS 39
A. PREPARATION OF DNA FROM VIRUS INFECTED CELLS 39
B. PREPARATION OF PLASMID DNA BY MODIFIED ALKALINE LYSIS 40
C. MEDIUM SCALE PREPARATION OF DNA FROM PLASMIDS FOR TRANSFECTION 40
D. POLYMERASE CHAIN REACTION 41
E. GEL ELECTROPHORESIS 41
F. PURIFICATION OF DNA BY COLUMN PURIFICATION AND RECOVERY OF DNA FROM
AGAROSE GELS 41
G. SEQUENCING 42
H. RESTRICTION FRAGMENT LENGTH POLYMORPHISM (RFLP) 42
I. CLONING METHODS 42
J. TWO-STEP EN PASSANT RED-MEDIATED RECOMBINATION 43
J.1. GENERAL PRINCIPLE FOR INSERTION OF SEQUENCES 44
J.2. GENERAL PRINCIPLE FOR DELETION OF SEQUENCES 46
K. PREPARATION OF COMPETENT CELLS AND TRANSFORMATION BY ELECTROPORATION
47
IMAGE 3
TABLE OF CONTENTS
I. SOUTHERN BLOTTING 48
M. SDS-PAGE 49
N. WESTERN BLOT 50
0. NEUTROPHIL ISOLATION FROM EQUINE BLOOD 51
P. CHEMOTAXIS ASSAYS 52
2.2.4. ANIMAL EXPERIMENTS 53
A. ANIMALS 53
B. ANIMAL CARE AND HOUSING 53
C. GENERAL PROCEDURES, INFECTION, HEALTH PARAMETERS 53
D. SACRIFICING ANIMALS 53
E. BAL AND TISSUE SAMPLING 53
F. FLOW CYTOMETRY 54
G. HISTOLOGY 54
H. TITRATION OF VIRUS FROM TISSUES 55
1 . STATISTICS 55
3. RESULTS 56
3.1. GENERATION OF MUTANT VIRUSES 56
3.1.1. DELETION OF GLYCOPROTEIN G IN EHV1 (VOH-C)3-AGG) 56
A. TRANSFER CONSTRUCTS 56
B. RECOMBINATION 57
3.1.2. INSERTION OF EHV-4 GG INTO EHV1 (VOH-03-4GG) 59
A. TRANSFER CONSTRUCT 59
B. RECOMBINATION 60
3.1.3. RE-INSERTION OF THE HYPERVARIABLE REGION OF EHV-1 INTO THE EHV-4
GG (VOH-034GG-HYP1) 61
A. TRANSFER CONSTRUCTS 61
B. RECOMBINATION 61
3.1.4. INSERTION OF EHV-4 GG HYPERVARIABLE REGION INTO EHV-1 GG
(VC)H-03-HYP4) 62
A. TRANSFER CONSTRUCTS 62
B. RECOMBINATION 62
3.1.5. RFLP OF RECOMBINANT BACS 63
3.1.6. RECONSTITUTION OF GP2 64
3.2. CHARACTERIZATION OF MUTANT VIRUSES 66
3.2.1. WESTERN BLOT 66
3.2.2. PLAQUE SIZE ASSAY 68
3.2.3. SINGLE-STEP GROWTH KINETICS 69
3.3. IN VITRO EXPERIMENTS 70
3.3.1. CHEMOTAXIS ASSAYS 70
3.4. IN VIVO EXPERIMENTS 72
IMAGE 4
TABLE OF CONTENTS
3.4.1. MOUSE EXPERIMENT I (PILOT EXPERIMENT) 72
A. BACK TITRATION OF VIRUS USED FOR INFECTION 72
B. CLINICAL EXAMINATION 72
C. WEIGHT LOSS MICE 72
D. VIRUS TITERS IN LUNGS 74
E. PATHOLOGY 75
3.4.2. MOUSE EXPERIMENT II 80
A. CLINICAL EXAMINATION 80
B. WEIGHT LOSS 80
C. VIRUS TITERS IN LUNGS 81
D. FACS ANALYSES 82
E. CYTOSPINS 87
4. DISCUSSION 88
4.1. CHARACTERIZATION OF MUTANT VIRUSES 88
4.2. IN VIVO EXPERIMENTS 91
5. REFERENCES 95
ACKNOWLEDGEMENTS I
I V
|
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| author | Thormann, Nora Mariella |
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| discipline | Agrar-/Forst-/Ernährungs-/Haushaltswissenschaft / Gartenbau |
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| isbn | 9783863870980 |
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| spelling | Thormann, Nora Mariella Verfasser (DE-588)1020595671 aut The role of the secreted glycoprotein G (gG) in equine herpesvirus type 1 (EHV-1) immune modulation and virulence vorgelegt von Nora Mariella Thormann Immunmodulation und Beeinflussung der Virulenz durch das Glykoprotein G (gG) des Equinen Herpesvirus vom Typ 1 Berlin Mensch und Buch Verl. 2011 [erschienen] 2012 VIII, 105, II S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Nebentitel: Immunmodulation und Beeinflussung der Virulenz durch das Glykoprotein G (gG) des Equinen Herpesvirus vom Typ 1 Berlin, Freie Univ., Diss., 2011 Equine herpesvirus 1 cabt Equine herpesvirus 4 cabt horse diseases cabt immune evasion cabt glycoproteins cabt neutrophils cabt chemotaxis cabt recombinatio cabt Equines Herpesvirus 1 (DE-588)4320759-5 gnd rswk-swf Pferdekrankheit (DE-588)4045515-4 gnd rswk-swf Virulenz (DE-588)4129834-2 gnd rswk-swf Glykoproteine (DE-588)4071905-4 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Pferdekrankheit (DE-588)4045515-4 s Equines Herpesvirus 1 (DE-588)4320759-5 s Virulenz (DE-588)4129834-2 s Glykoproteine (DE-588)4071905-4 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:kobv:188-fudissthesis000000036272-5 http://www.diss.fu-berlin.de/diss/receive/FUDISS_thesis_000000036272 kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024822250&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Thormann, Nora Mariella The role of the secreted glycoprotein G (gG) in equine herpesvirus type 1 (EHV-1) immune modulation and virulence Equine herpesvirus 1 cabt Equine herpesvirus 4 cabt horse diseases cabt immune evasion cabt glycoproteins cabt neutrophils cabt chemotaxis cabt recombinatio cabt Equines Herpesvirus 1 (DE-588)4320759-5 gnd Pferdekrankheit (DE-588)4045515-4 gnd Virulenz (DE-588)4129834-2 gnd Glykoproteine (DE-588)4071905-4 gnd |
| subject_GND | (DE-588)4320759-5 (DE-588)4045515-4 (DE-588)4129834-2 (DE-588)4071905-4 (DE-588)4113937-9 |
| title | The role of the secreted glycoprotein G (gG) in equine herpesvirus type 1 (EHV-1) immune modulation and virulence |
| title_alt | Immunmodulation und Beeinflussung der Virulenz durch das Glykoprotein G (gG) des Equinen Herpesvirus vom Typ 1 |
| title_auth | The role of the secreted glycoprotein G (gG) in equine herpesvirus type 1 (EHV-1) immune modulation and virulence |
| title_exact_search | The role of the secreted glycoprotein G (gG) in equine herpesvirus type 1 (EHV-1) immune modulation and virulence |
| title_full | The role of the secreted glycoprotein G (gG) in equine herpesvirus type 1 (EHV-1) immune modulation and virulence vorgelegt von Nora Mariella Thormann |
| title_fullStr | The role of the secreted glycoprotein G (gG) in equine herpesvirus type 1 (EHV-1) immune modulation and virulence vorgelegt von Nora Mariella Thormann |
| title_full_unstemmed | The role of the secreted glycoprotein G (gG) in equine herpesvirus type 1 (EHV-1) immune modulation and virulence vorgelegt von Nora Mariella Thormann |
| title_short | The role of the secreted glycoprotein G (gG) in equine herpesvirus type 1 (EHV-1) immune modulation and virulence |
| title_sort | the role of the secreted glycoprotein g gg in equine herpesvirus type 1 ehv 1 immune modulation and virulence |
| topic | Equine herpesvirus 1 cabt Equine herpesvirus 4 cabt horse diseases cabt immune evasion cabt glycoproteins cabt neutrophils cabt chemotaxis cabt recombinatio cabt Equines Herpesvirus 1 (DE-588)4320759-5 gnd Pferdekrankheit (DE-588)4045515-4 gnd Virulenz (DE-588)4129834-2 gnd Glykoproteine (DE-588)4071905-4 gnd |
| topic_facet | Equine herpesvirus 1 Equine herpesvirus 4 horse diseases immune evasion glycoproteins neutrophils chemotaxis recombinatio Equines Herpesvirus 1 Pferdekrankheit Virulenz Glykoproteine Hochschulschrift |
| url | http://www.diss.fu-berlin.de/diss/receive/FUDISS_thesis_000000036272 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024822250&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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