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Language(s):
eng - English
Dates:
2006-12
Publication Status:
Accepted / In Press
Pages:
65 p.
Publishing info:
Kiel : Christian-Albrechts-Universität
Table of Contents:
1. SUMMARY .................................................................. 5
2. DEUTSCHE ZUSAMMENFASSUNG................................ 7
3. INTRODUCTION......................................................... 9
3.1. MAJOR HISTOCOMPATIBILITY COMPLEX................................................9
3.2. REAL-TIME PCR..................................................................................10
3.3. THESIS OUTLINE ..................................................................................11
4. MATERIAL AND METHODS........................................ 15
4.1. FISH FOR THE INFECTION EXPERIMENT ................................................15
4.2. TREATMENT OF CELL CULTURES .........................................................17
4.3. PRIMER DESIGN ...................................................................................19
4.4. SEQUENCING OF GENES .......................................................................20
4.5. RNA EXTRACTION AND REVERSE TRANSCRIPTION ..............................22
4.6. REAL-TIME PCR..................................................................................23
4.7. HOUSEKEEPING GENES ........................................................................24
4.8. DATA ANALYSIS AND STATISTICAL METHODS .....................................26
5. RESULTS .................................................................. 27
5.1. EVALUATION OF PCR CONDITIONS AND PRIMERS ...............................27
5.2. SELECTION OF HOUSEKEEPING GENES IN CELL CULTURES ...................29
5.2.1. Cultures of head kidney cells ...........................................................29
5.2.2. Cultures of spleen cells ....................................................................31
5.3. SELECTION OF HOUSEKEEPING GENES IN FISH......................................33
5.3.1. Expression in head kidneys ..............................................................33
5.3.2. Expression in spleens .......................................................................34
5.3.3. Expression in gills ............................................................................36
5.4. CELL CULTURES ..................................................................................38
5.4.1. MHC II-expression in cell cultures ..................................................38
5.4.2. Number of cells ................................................................................40
5.5 INFECTION EXPERIMENT......................................................................41
5.5.1. Immune response genes in spleens...................................................41
5.5.2 Immune response genes in head kidneys...........................................44
5.5.3 Immune response genes in gills.........................................................46
6. DISCUSSION............................................................ 49
6.1. PRIMER DESIGN AND PCR ...................................................................49
6.2. HOUSEKEEPING GENE..........................................................................51
6.3. CELL CULTURE EXPERIMENT ...............................................................53
6.4. INFECTION EXPERIMENT......................................................................56
7. LITERATURE ............................................................ 59
8. DANKSAGUNG.......................................................... 63
STELLUNGNAHME ........................................................ 65
Rev. Type:
-
Identifiers:
eDoc: 294089
Other: Dipl/11347
Degree:
Diploma