Apoptosis regulation by Raf-1 and Bcl-2:
Saved in:
| Main Author: | |
|---|---|
| Format: | Book |
| Language: | English German |
| Published: |
1999
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Würzburg, Univ., Diss., 2000. - Zsfassung in dt. Sprache |
| Physical Description: | 84 Bl. Ill., graph. Darst. |
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Record in the Search Index
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| adam_text |
CONTENTS
SUMMARY ^
CHAPTER 1. INTRODUCTION 13
1-1. APOPTOSIS 13
1-2. THE BCL-2 FAMILY MEMBERS IN APOPTOSIS REGULATION 17
1-3. RAF-1 AND ITS ISOFORMS A-, AND B-RAF 19
1- 4. THE AIM OF THIS STUDY 21
CHAPTER 2. MATERIALS AND METHODS 23
2- 1. MATERIALS 23
2-1-1 CHEMICALS AND ENZYMES 23
2-1-2 ISOTOPE 25
2-1 -3. MOLECULAR WEIGHT MARKERS 25
2-1-4. BUFFERS 26
2-1-5. KITS 30
2-1-6. ANTIBODIES 31
2-1-7. PLASMIDS 32
2-1-8. VIRUSES 32
2-1-9. PCR-PRIMERS 32
5
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/959863656
2-1-10. OTHERS
33
2-1-11. ANIMALS 33
2-2 METHODS 34
2-2-1. ESTABLISHMENT OF PRIMARY CELL CULTURE 34
2-2-1 -1. ISOLATION OF EMBRYOS 34
2-2-1 -2. CELL LINES AND CULTURE CONDITIONS 34
2-2-1-3. ISOLATION OF GENOMIC DNA FROM EMBRYONIC TISSUES 35
2-2-1-4. GENOTYPING OF
BCL-2 DEFICIENT CELLS 35
2-2-1-5. GENOTYPING OF
RAF-L DEFICIENT CELLS 35
2-2-2. RETROVIRAL INFECTION 36
2-2-2-1. HARVEST AND TITRATION OF ONCOGEN CONTAINING VIRUSES 36
2-2-2-2. INFECTION 37
2-2-3. DNA TRANSFECTION 37
2-2-3-1. PLASMID CONSTRUCTS 37
2-2-3-2. AMPLIFICATION OF PLASMID DNA 37
2-2-3-3. PLASMID ISOLATION USING QIAGEN PROTOCOL 38
2-2-3-4 THE MEASUREMENT OF PLASMID CONCENTRATION 38
2-2-3-5. TRANSFECTION OF ADHERENT CELLS 39
2-2-4. DNA RELEASE ASSAY 39
2-2-5. ANALYSIS OF DNA FRAGMENTATION BY AGAROSE GEL ELECTROPHORESIS 40
2-2-6. THE MEASUREMENT OF PROTEIN CONCENTRATION 40
2-2-7. IMMUNOPRECIPITATION 41
6
2-2-8. WESTERN BLOT ANALYSIS 41
2-2-9 ANTISENSE EXPERIMENTS 41
2-2-10 RT-PCR OF
BAD 42
2-2-10-1. TOTAL RNA EXTRACTION USING THE Q1AGEN RNEASY MIDI KIT 42
2-2-10-2. CDNA SYNTHESIS USING 1ST STRAND CDNA SYNTHESIS KIT
FOR RT-PCR (AMV) 43
2- 2-10-3. RT-PCR 44
CHAPTER 3. RESULTS 45
3- 1. APOPTOSIS SUPPRESSION BY MITOCHONDRIA TARGETED ONCOGENIC
RAF-1 DOES NOT REQUIRE BCL-2 45
3-1-1. THE ESTABLISHMENT OF
BCL-2 DEFICIENT CELLS 45
3-1-2.
BCL-2 DEFICIENT FIBROBLASTS SHOW INCREASED SENSITIVITY TOWARDS
DIFFERENT CLASSES OF APOPTOTIC STIMULI 48
3-1 -3. OVEREXPRESSION OF BCL-2 INHIBITS DOXORUBICIN-INDUCED APOPTOSIS
49
3-1-4. BCL-2 BLOCKS CASPASE-3 ACTIVATION 50
3-1-5. GAG-V-RAF PROTEAS
BCL-2 DEFICIENT CELLS FROM APOPTOSIS
BY BLOCKING CASPASE-3 ACTIVATION 52
3-1-6. ACTIVATED RAF-1 REQUIRES MITOCHONDRIAL TRANSLOCATION
FOR APOPTOSIS SUPPRESSION 54
3-1-7. ABLATION OF RAF-1 EXPRESSION DOES NOT AFFEA DOXORUBICIN-INDUCED
CELL DEATH IN
BCL-2 DEFICIENT AND WILD TYPE CELLS 58
7
! 3-2. BCL-2 INHIBITS DOXOTUBICIN-INDUCED APOPTOSIS IN
RAF-1 DEFICIENT FIBROBLASTS 60
3-2-1. THE ESTABLISHMENT OF
RAF-1 DEFICIENT CELLS 60
3-2-2. THE
RAF-1 DEFICIENT CELLS SHOW INCREASED SENSITIVITY TOWARDS
DOXORUBICIN-INDUCED APOPTOSIS 61
3-2-3. THE OVEREXPRESSION OF BCL-2 INHIBITS DOXORUBICIN-INDUCED
APOPTOSIS
IN
RAF-1 AND WILD TYPE CELLS 64
CHAPTER 4. DISCUSSION 67
CHAPTER 5. REFERENCES 73
CHAPTER 6. ABBREVIATIONS 83
8 |
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| physical | 84 Bl. Ill., graph. Darst. |
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| spelling | Zhong, Jian 1961- Verfasser (DE-588)122373464 aut Apoptosis regulation by Raf-1 and Bcl-2 von Jian Zhong 1999 84 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Würzburg, Univ., Diss., 2000. - Zsfassung in dt. Sprache Hochschulschrift gtt Apoptosis (DE-588)4291096-1 gnd rswk-swf Regulation (DE-588)4049075-0 gnd rswk-swf Mikroform (DE-588)4039216-8 gnd rswk-swf Molekularbiologie (DE-588)4039983-7 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Apoptosis (DE-588)4291096-1 s Regulation (DE-588)4049075-0 s Molekularbiologie (DE-588)4039983-7 s Mikroform (DE-588)4039216-8 s 1\p DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008921861&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk |
| spellingShingle | Zhong, Jian 1961- Apoptosis regulation by Raf-1 and Bcl-2 Hochschulschrift gtt Apoptosis (DE-588)4291096-1 gnd Regulation (DE-588)4049075-0 gnd Mikroform (DE-588)4039216-8 gnd Molekularbiologie (DE-588)4039983-7 gnd |
| subject_GND | (DE-588)4291096-1 (DE-588)4049075-0 (DE-588)4039216-8 (DE-588)4039983-7 (DE-588)4113937-9 |
| title | Apoptosis regulation by Raf-1 and Bcl-2 |
| title_auth | Apoptosis regulation by Raf-1 and Bcl-2 |
| title_exact_search | Apoptosis regulation by Raf-1 and Bcl-2 |
| title_full | Apoptosis regulation by Raf-1 and Bcl-2 von Jian Zhong |
| title_fullStr | Apoptosis regulation by Raf-1 and Bcl-2 von Jian Zhong |
| title_full_unstemmed | Apoptosis regulation by Raf-1 and Bcl-2 von Jian Zhong |
| title_short | Apoptosis regulation by Raf-1 and Bcl-2 |
| title_sort | apoptosis regulation by raf 1 and bcl 2 |
| topic | Hochschulschrift gtt Apoptosis (DE-588)4291096-1 gnd Regulation (DE-588)4049075-0 gnd Mikroform (DE-588)4039216-8 gnd Molekularbiologie (DE-588)4039983-7 gnd |
| topic_facet | Hochschulschrift Apoptosis Regulation Mikroform Molekularbiologie |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008921861&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT zhongjian apoptosisregulationbyraf1andbcl2 |