Interference of Hepatitis C virus (HCV) core protein with intracellular signal transduction processes in liver cells:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
Aachen
Shaker
2001
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| Schriftenreihe: | Berichte aus der Biologie
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Zugl.: Düsseldorf, Univ., Diss., 2000 |
| Beschreibung: | V, 128 S. Ill., graph. Darst. |
| ISBN: | 3826587685 |
Internformat
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Datensatz im Suchindex
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| adam_text | Table of Contents
Table of Contents I
Abbreviations IV
1. Introduction 1
1.1 Historical review 1
1.2 Pathogenesis and pathology of hepatitis C virus 2
1.3 Characteristics of the virion 6
1.4 Classification and Genome Organization 7
1.5 Hepatitis C virus core protein 11
1.6 Signal transduction processes 13
1.7 Tetracycline regulated system 17
1.8 The scope of this thesis 20
2. Material and Methods 23
2.1 Materials 23
2.1.1 Chemicals, Enzymes, and Antibodies 23
2.1.2 Vectors and bacterial strains 23
2.1.3 Cell lines 23
2.1.4 Media for cell culture 23
2.1.5 Chemicals and Enzymes 23
2.1.6 Antibodies 24
2.1.7 Kits for molecular biology 25
2.1.8 Membranes for RNA and protein blotting 25
2.1.9 X ray films 25
2.1.10 Media for bacteria 25
2.1.11 Buffers and solutions for DNA electrophoresis 26
2.1.12 Buffers and solutions for DNA sequencing 26
2.1.13 Buffers and solutions for RNA electrophoresis and Northern blot 27
2.1.14 Buffers and solutions for the preparation of nuclear and whole cell 28
extracts
2.1.15 Buffers and solutions for protein electrophoresis and Western blot 29
2.1.16 Buffers and solutions for eiectrophoretic mobility shift assay (EMSA) 31
i
2.1.17 Buffers and solutions for protein kinase assays 32
2.2. Methods 33
2.2.1 Reverse Transcriptase Polymerase Chain Reaction (RT PCR) 33
2.2.2 Cloning of PCR products in eukaryotic expression vector 33
2.2.3 Transformation of E.Coli with the constructed plasmid 33
2.2.4 Isolation of plasmid DNA 34
2.2.5 Sequencing of plasmid DNA 34
2.2.6 Extraction of DNA fragments from agrose gel 35
2.2.7 Generation of HCV core (160 and 191) and plasmid construction 35
2.2.8 Development of double stable Tet Off liver cell line 36
2.2.8.2 Screening of Tet Off stable cell lines 36
2.2.8.4 Screening of double stable Tet Off cell line 37
2.2.8.5 Luciferase assay 37
2.2.9 Measurement of cell viability and proliferation 37
2.2.10 Measurement of cytotoxicity 38
2.2.11 Northern blot 38
2.2.11.1 Isolation of total RNAfrom cultured cells 38
2.2.11.2 RNA gel electrophoresis 39
2.2.11.3 Transfer of fractionated RNA from formaldehyde gel to membrane 39
2.2.11.4 Labelling DNA probes by Random primer extension 40
2.2.11.5 Purification of labelled DNA probes 40
2.2.11.6 Hybridization of the immobilized RNA to a sequence specific probe 40
2.2.12.1 Protein gel electrophoresis (SDS PAGE) 41
2.2.12.2 Protein transfer and detection of the specific signal 41
2.2.13 Electrophoretic mobility shift assay (EMSA) 41
2.2.13.1 Preparation of nuclear extracts 42
2.2.13.2 End labelling of oligonucleotides 42
2.2.13.3 Removal of unincorporated label 42
2.2.13.4 DNA binding reaction using nuclear extract 42
2.2.13.5 Electrophoresis of DNA protein complexes 43
2.2.14 Protein kinase assays 43
ii
3. Results 44
3.1 Generation of the HCV core genomic regions 44
3.2 Development and screening of Tet Off liver cell line 47
3.3 Development and Screening of double stable Tet Off liver cell line 47
3.4 Tightly regulated expression of HCV core protein 50
3.5 Characterization of viral cell interactions of HCV core and liver cells 53
3.5.1 Measurement of cytotoxicity 54
3.5.2 Measurement of cell proliferation 55
3.5.3 Effect of HCV core protein on cell morphology 57
3.5.4 Effect of HCV core protein on proto oncogenes and tumor 59
suppressor genes
3.5.5 Effect of HCV core protein on MAPK and MAK phosphatase 1 71
3.5.6 Effect of HCV core protein on transcription factors 78
3.5.7 Effect of HCV core protein on apoptosis related genes 86
3.5.8 Effect of HCV core protein on pro and anti apoptotic genes 88
3.5.9 Effect of HCV core protein on HSP70 90
4. Discussion 91
4.1 Tetracycline regulated system in HepG2 cells 91
4.2 Interaction of HCV core protein with intracellular signal transduction 92
processes in liver cells
4.2.1 Influence of HCV core protein on cell proliferation and 95
transcription factors
4.2.2 Influence of HCV core protein on MAP 1 and MAP kinases 99
4.3 Role of HCV core protein in the development of HCC 101
4.4 Role of HCV core protein in apoptosis 104
5.Summery 109
6.Zusammenfassung 110
7.References 111
8.Appendix 130
ni
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| indexdate | 2024-07-09T20:02:32Z |
| institution | BVB |
| isbn | 3826587685 |
| language | English |
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| physical | V, 128 S. Ill., graph. Darst. |
| publishDate | 2001 |
| publishDateSearch | 2001 |
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| publisher | Shaker |
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| series2 | Berichte aus der Biologie |
| spelling | Hassan, Mohamed Verfasser aut Interference of Hepatitis C virus (HCV) core protein with intracellular signal transduction processes in liver cells Mohamed Hassan Aachen Shaker 2001 V, 128 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Berichte aus der Biologie Zugl.: Düsseldorf, Univ., Diss., 2000 Signaltransduktion (DE-588)4318717-1 gnd rswk-swf Genexpression (DE-588)4020136-3 gnd rswk-swf Hepatitis-C-Virus (DE-588)4262008-9 gnd rswk-swf Proteine (DE-588)4076388-2 gnd rswk-swf Interferenz Virologie (DE-588)4329145-4 gnd rswk-swf Core (DE-588)4344922-0 gnd rswk-swf Leberepithelzelle (DE-588)4120659-9 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Leberepithelzelle (DE-588)4120659-9 s Hepatitis-C-Virus (DE-588)4262008-9 s Core (DE-588)4344922-0 s Proteine (DE-588)4076388-2 s Genexpression (DE-588)4020136-3 s DE-604 Interferenz Virologie (DE-588)4329145-4 s Signaltransduktion (DE-588)4318717-1 s HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=012995533&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Hassan, Mohamed Interference of Hepatitis C virus (HCV) core protein with intracellular signal transduction processes in liver cells Signaltransduktion (DE-588)4318717-1 gnd Genexpression (DE-588)4020136-3 gnd Hepatitis-C-Virus (DE-588)4262008-9 gnd Proteine (DE-588)4076388-2 gnd Interferenz Virologie (DE-588)4329145-4 gnd Core (DE-588)4344922-0 gnd Leberepithelzelle (DE-588)4120659-9 gnd |
| subject_GND | (DE-588)4318717-1 (DE-588)4020136-3 (DE-588)4262008-9 (DE-588)4076388-2 (DE-588)4329145-4 (DE-588)4344922-0 (DE-588)4120659-9 (DE-588)4113937-9 |
| title | Interference of Hepatitis C virus (HCV) core protein with intracellular signal transduction processes in liver cells |
| title_auth | Interference of Hepatitis C virus (HCV) core protein with intracellular signal transduction processes in liver cells |
| title_exact_search | Interference of Hepatitis C virus (HCV) core protein with intracellular signal transduction processes in liver cells |
| title_full | Interference of Hepatitis C virus (HCV) core protein with intracellular signal transduction processes in liver cells Mohamed Hassan |
| title_fullStr | Interference of Hepatitis C virus (HCV) core protein with intracellular signal transduction processes in liver cells Mohamed Hassan |
| title_full_unstemmed | Interference of Hepatitis C virus (HCV) core protein with intracellular signal transduction processes in liver cells Mohamed Hassan |
| title_short | Interference of Hepatitis C virus (HCV) core protein with intracellular signal transduction processes in liver cells |
| title_sort | interference of hepatitis c virus hcv core protein with intracellular signal transduction processes in liver cells |
| topic | Signaltransduktion (DE-588)4318717-1 gnd Genexpression (DE-588)4020136-3 gnd Hepatitis-C-Virus (DE-588)4262008-9 gnd Proteine (DE-588)4076388-2 gnd Interferenz Virologie (DE-588)4329145-4 gnd Core (DE-588)4344922-0 gnd Leberepithelzelle (DE-588)4120659-9 gnd |
| topic_facet | Signaltransduktion Genexpression Hepatitis-C-Virus Proteine Interferenz Virologie Core Leberepithelzelle Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=012995533&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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