Identification and functional characterization of MiRNAs in gammaherpesvirus infection:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2010
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| Schlagworte: | |
| Online-Zugang: | Volltext http://d-nb.info/1008071315/34 https://nbn-resolving.org/urn:nbn:de:bvb:19-121726 Inhaltsverzeichnis |
| Beschreibung: | 140 Bl. graph. Darst. |
Internformat
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Datensatz im Suchindex
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| adam_text | IMAGE 1
DISSERTATION ZUR ERLANGUNG DES DOKTORGRADES
DER FAKULTAET FUER CHEMIE UND PHARMAZIE DER LUDWIG-MAXIMILIAN-UNIVERSITAET
MUENCHEN
IDENTIFICATION AND FUNCTIONAL CHARACTERIZATION OF
MIRNAS IN GAMMAHERPESVIRUS INFECTION
JIAYUN ZHU
AUS
SHANGHAI, CHINA
2010
BIBLIOGRAFISCHE INFORMATIONEN HTTP://D-NB.INFO/1008109975
DIGITALISIERT DURCH
IMAGE 2
SUMMARY 1
1. INTRODUCTION 1
1.1 MIRNAGENES 1
1.1.1 MIRNAS AND AGO PROTEINS 1
1.1.2 IDENTIFICATION OFMIRNA GENES AND MIRNA DATABASE 3
1.1.3 MIRNAS AND CANCERS 4
1.2 GAMMAHERPESVIRUS AND MIRNAS 5
1.2.1 EPSTEIN-BARR VIRUS AND MURINE HERPESVIRUS 68 5
1.2.2 IDENTIFICATION OFMIRNAS IN GAMMAHERPESVIRUSES 7
1.2.3 MIRNA FUNCTIONS IN GAMMAHERPESVIRUS INFECTION 8
1.3 NEXT GENERATION SEQUENCING AND MIRNA STUDY 9
1.3.1 ADVENT OF NEXT GENERATION SEQUENCING 9
1.3.2 WORKING PRINCIPLES OF 454 SEQUENCING 9
1.3.3 WORKING PRINCIPLES OFSOLEXA SEQUENCING 12
1.3.4 SMALL RNA SEQUENCING BY NGS 12
1.3.5 ANALYSIS TOOLS FOR NGS DATA 14
1.4 A IM OF THE THESIS 15
2. RESULT 16
2.1 NGS DATA PROCESSING SOFTWARE 16
2.2 ANALYSIS OF SMALL RNA LIBRARIES FROM EBV-INFECTED NPC SAMPLES 18
2.2.1 SMALL RNA CLONING AND SEQUENCING FROM NPC AND CONTROL SAMPLES 18
2.2.2 IDENTIFICATION OF NOVEL EBV MIRNA GENES 19
2.2.3 IDENTIFICATION OF NOVEL HUMAN MIRNA GENES 21
2.2.4 VALIDATION OF NOVEL EBV MIRNAS 22
2.2.5 MIRNA EXPRESSION ANALYSIS SUGGESTS AN NPC-SPECIFIC MIRNA
SIGNATURE. 24
2.2.6 MISREGULATED MIRNAS HSA-MIR-15A AND HSA-MIR-16 INHIBIT EXPRESSION
OF TUMOR SUPPRESSOR
BRCA1 26
2.2.7 SUMMARY OF RESULTS FROM NGS DATA IN NPC SAMPLES 28
2.3 ANALYSIS OF SMALL RNA LIBRARIES FROM MHV-68 INFECTED CELL LINES 28
2.3.1 SMALL RNA CLONING AND SEQUENCING OF MHV-68 INFECTED CELLS 29
2.3.2 IDENTIFICATION OF NOVEL MHV-68 MIRNA GENES 30
2.3.3 GENOMIC ORGANIZATION OF NOVEL MHV-68 MIRNAS 33
2.3.4 VALIDATION OF THE NOVEL MHV-68 MIRNA EXPRESSION 35
2.3.5 CELLULAR MIRNA PROFILES IN NIH3T3 AND SLL CELLS 37
2.3.6 SUMMARY OF RESULTS FROM NGS DATA IN MHV-68 INFECTED CELLS 41
IMAGE 3
2.4 MIS-ANNOTATION OF MICRORNA GENES IN MIRBASE 41
2.4.1 EXAMINATION OFMURINE MIRNA GENES 42
2.4.2 EXAMINATION OF HUMAN MIRNA GENES 47
2.4.3 SUMMARY OF EXAMINATION OF MIS-ANNOTATED MIRNAS IN MIRBASE 50
3. DISCUSSION 52
3.1 ADVANTAGES OF DEVELOPED SOFTWARE FOR NGS DATA ANALYSIS 52
3.2 IDENTIFICATION OF NOVEL GAMMAHERPESVIRUS MIRNAS 54
3.3 GENOME ORGANIZATION AND EVOLUTIONARY CONSERVATION OF
GAMMAHERPESVIRUS MLRNAS 55
3.4 EBVANDTUMORIGENESIS 57
3 .5 M H V - 68 M I R NA EXPRESSION SIGNATURES 59
3.6 FURTHER APPLICATION OF NGS IN M I R NA STUDIES 60
4. MATERIALS & METHODS 63
4.1 MATERIALS 63
4.1.1 CHEMICALS AND ENZYMES 63
4.1.2 PLASMID. 63
4.1.3 ANTIBODIES 63
4.1.4 TISSUE SAMPLES 64
4.1.5 BACTERIAL STRAINS, VIRUS STOCK AND CELL LINES 64
4.1.6 CELL CULTURE MEDIA 65
4.1.7 BUFFERS AND SOLUTIONS 66
4.2 METHODS 70
4.2.1 MOLECULAR BIOLOGICAL METHODS 70
4.2.1.1 GENERAL METHODS 70
4.2.1.2 CLONING OF 3 UTRS FROM CDNA LIBRARY INTO PMIR-TK-RNL 70
4.2.1.3 RNA EXTRACTION FROM TISSUE SAMPLES OR CULTURED CELLS 70
4.2.1.4 AGO COMPLEX PURIFICATION 71
4.2.1.5 RNA POLYACRYLAMIDE GEL ELECTROPHORESIS 72
4.2.1.6 NORTHERN BLOTTING 72
4.2.1.7 REVERSE TRANSCRIPTION AND QRT-PCR OF MRNA DETECTION 73
4.2.1.8 REVERSE TRANSCRIPTION AND QRT-PCR OF MIRNA DETECTION 74
4.2.1.9 REVERSE TRANSCRIPTION AND STEM-LOOP QRT-PCR OF MIRNA DETECTION
75
4.2.1.10 WESTERN BLOTTING 75
4.2.1.11 GENERATION OF SMALL RNA LIBRARIES FOR 454 SEQUENCING 76
4.2.2 CELLULAR BIOLOGICAL METHODS 77
4.2.2.1 CULTURING OF MAMMALIAN CELLS 77
4.2.2.2 VIRUS INFECTION AND LYTIC CYCLE INDUCTION 77
4.2.2.3 2 0ME- MIRNA INHIBITOR TRANSFECTION 77
4.2.2.4 COTRANSFECTION OF 2 OME- INHIBITORS AND LUCIFERASE REPORTER
CONSTRUCTS 78
IMAGE 4
4.2.2.5 LUCIFERASE ASSAY 78
4.2.3 COMPUTATIONAL METHODS 78
4.2.3.1 SEQUENCE ANALYSIS OF 454 SEQUENCING LIBRARIES 78
4.2.3.2 PROGRAMMING METHOD 79
ABBREVIATIONS 80
FIGURE INDEX 82
TABLE INDEX 83
REFERENCES 84
APPENDIX 92
APPENDIX TABLE I 92
APPENDIX TABLE II 100
APPENDIX TABLE III 112
APPENDIX TABLE IV 125
PUBLICATIONS 136
ACKNOWLEDGEMENTS 137
CURRICULUM VITAE 139
|
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| spelling | Zhu, Jiayun 1981- Verfasser (DE-588)142706167 aut Identification and functional characterization of MiRNAs in gammaherpesvirus infection Jiayun Zhu 2010 140 Bl. graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Univ., Diss., 2010 (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:19-121726 PDF http://edoc.ub.uni-muenchen.de/12172/ Verlag kostenfrei Volltext http://d-nb.info/1008071315/34 Langzeitarchivierung Nationalbibliothek https://nbn-resolving.org/urn:nbn:de:bvb:19-121726 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=020766750&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Zhu, Jiayun 1981- Identification and functional characterization of MiRNAs in gammaherpesvirus infection |
| subject_GND | (DE-588)4113937-9 |
| title | Identification and functional characterization of MiRNAs in gammaherpesvirus infection |
| title_auth | Identification and functional characterization of MiRNAs in gammaherpesvirus infection |
| title_exact_search | Identification and functional characterization of MiRNAs in gammaherpesvirus infection |
| title_full | Identification and functional characterization of MiRNAs in gammaherpesvirus infection Jiayun Zhu |
| title_fullStr | Identification and functional characterization of MiRNAs in gammaherpesvirus infection Jiayun Zhu |
| title_full_unstemmed | Identification and functional characterization of MiRNAs in gammaherpesvirus infection Jiayun Zhu |
| title_short | Identification and functional characterization of MiRNAs in gammaherpesvirus infection |
| title_sort | identification and functional characterization of mirnas in gammaherpesvirus infection |
| topic_facet | Hochschulschrift |
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