The synaptonemal complex and its role in crossover regulation during C. elegans meiosis
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| 015 | |a 24,H01 |2 dnb | ||
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| 100 | 1 | |a Neves, Ana Rita Rodrigues |d 1994- |e Verfasser |0 (DE-588)1310873097 |4 aut | |
| 245 | 1 | 0 | |a The synaptonemal complex and its role in crossover regulation during C. elegans meiosis |c presented by M.Sc. Ana Rita Rodrigues Neves ; referees: Dr. Sara Cuylen-Häring, Prof. Dr. Michael Knop |
| 264 | 1 | |a Heidelberg |c [2023] | |
| 300 | |a xxii, 219 Seiten |b Illustrationen, Diagramme |c 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Ruprecht-Karls-University Heidelberg |d 2023 | ||
| 546 | |a Text Englisch, Zusammenfassung in englischer und deutscher Sprache | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 700 | 1 | |a Cuylen, Sara |0 (DE-588)1024182436 |4 dgs | |
| 710 | 2 | |a Universität Heidelberg |0 (DE-588)2024349-2 |4 dgg | |
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CONTENTS
ACKNOWLEDGEMENTS
.
III
SUMMARY
.
V
ZUSAMMENFASSUNG
.
VII
LIST
OF
FIGURES
.
XVIII
LIST
OF
TABLES
.
XX
LIST
OF
ABBREVIATIONS
.
XXI
CHAPTER
1
INTRODUCTION
1
1.1
MEIOSIS
.
1
1.2
MEIOTIC
PROPHASE
I
IN
C.
ELEGANS
.
2
1.3
THE
SYNAPTONEMAL
COMPLEX
.
4
1.3.1
ORGANISATION
OF
THE
SYNAPTONEMAL
COMPLEX
.
6
1.3.2
SYNAPSIS
INITIATION
AND
ELONGATION
.
11
1.3.3
POST-TRANSLATIONAL
MODIFICATION
AND
THE
SYNAPTONEMAL
COMPLEX
.
.
12
1.4
CROSSOVER
REGULATION
.
13
1.4.1
THE
SYNAPTONEMAL
COMPLEX
AND
CROSSOVER
REGULATION
.
16
1.4.2
MODELS
OF
CROSSOVER
INTERFERENCE
.
17
CHAPTER
2
AIMS
21
CHAPTER
3
MATERIALS
AND
METHODS
23
3.1
MAINTENANCE
AND
CULTURE
CONDITIONS
OF
C.
ELEGANS
STRAINS
.
23
3.2
GENERATION
OF
C.
ELEGANS
STRAINS
BY
CRISPR/CAS9-MEDIATED
GENOME
EDITING
25
3.3
VIABILITY
ASSAYS
.
26
3.4
SYNCHRONISATION
OF
C.
ELEGANS
ANIMALS
BY
BLEACHING
.
26
3.5
ISOLATION
OF
MEIOTIC
NUCLEI
FROM
A
SYNCHRONISED
POPULATION
OF
C.
ELEGANS
.
27
3.6
PREPARATION
OF
WHOLE
WORM
AND
MEIOTIC
NUCLEI
LYSATES
.
29
3.7
SDS-PAGE
AND
IMMUNOBLOTTING
.
30
3.8
SILVER
STAINING
.
31
3.9
2C
METHOD:
A
SILICA-BASED
SOLID-PHASE
EXTRACTION
OF
CROSSLINKED
NUCLEIC
ACID-BOUND
PROTEINS
.
32
3.10
ECLIP-SEQ:
ENHANCED
CROSSLINKING
AND
IMMUNOPRECIPITATION
FOLLOWED
BY
SEQUENCING
.
33
3.10.1
IMMUNOPRECIPITATION
.
33
3.10.2
RNA
EXTRACTION
AND
3
'
RNA
ADAPTER
LIGATION
.
35
3.10.3
CDNA
LIBRARY
PREPARATION
.
37
3.10.4
HIGH
THROUGHPUT
SEQUENCING
AND
ANALYSIS
.
38
3.11
POLYNUCLEOTIDE KINASE
(PNK)
ASSAY
.
40
3.12
PROTEOMIC
ANALYSIS
OF
SYP-4
.
41
3.12.1
PHOSPHOPROTEOMICS
.
41
3.12.2
IDENTIFICATION
OF
S
YP-4-INTERACTING PROTEINS
BY
MASS
SPECTROMETRY
44
3.13
QUANTIFICATION
OF
SYP-4
PROTEIN
LEVELS
.
46
3.14
IMMUNOFLUORESCENCE
.
46
3.15
FLUORESCENCE
MICROSCOPY
.
48
3.16
QUANTIFICATION
OF
TRANSITION
ZONE
.
49
3.17
QUANTIFICATION
OF
SYP
LEVELS
DURING
PROPHASE
I
.
49
3.18
QUANTIFICATION
OF
NUMBER
OF
COSA
1
FOCI
AND
DAPI-STAINING
BODIES
.
.
.
51
3.19
GENETIC
MAPPING
OF
RECOMBINATION
SITES
.
52
3.20
CROSSOVER
INTERFERENCE
ANALYSIS
.
57
3.21
STATISTICS
.
58
3.22
SYP-4
MULTIPLE
SEQUENCE
ALIGNMENT,
AMINO
ACID
CONSERVATION
AND
DISOR
DER
PREDICTION
.
59
3.23
YEAST
TWO-HYBRID
.
59
3.23.1
PREPARATION
OF
TWO-HYBRID
CONSTRUCTS
.
60
3.23.2
TRANSFORMATION
OF
YEAST
CELLS
.
60
3.23.3
MATING
OF
YEAST
CELLS
.
61
3.23.4
ASSAYING
^-GALACTOSIDASE
ACTIVITY
.
62
CHAPTER
4
IDENTIFICATION
OF
RNAS
WITHIN
THE
SYNAPTONEMAL
COMPLEX
63
4.1
IDENTIFICATION
OF
RNA-INTERACTING
SYNAPTONEMAL
COMPLEX
COMPONENTS
.
.
64
4.2
IDENTIFICATION
OF
HIM-3-BOUND
RNAS
BY
ECLIP-SEQ
.
66
4.3
QUANTIFICATION
OF
SYNAPSIS
DEFECTS
IN
THE
ABSENCE
OF
A
PUTATIVE
HIM-3
INTERACTING
RNA
.
68
4.4
DISCUSSION
.
71
CHAPTER
5
UNDERSTANDING
THE
ROLE
OF
THE
SYNAPTONEMAL
COMPLEX
IN
CROSSOVER
REGULATION
75
5.1
CHARACTERISATION
OF
MEIOTIC
DEFECTS
IN
C-TERMINALLY
TRUNCATED
SYP-4
ANI
MALS
(A114)
.
75
5.2
IDENTIFICATION
OF
PUTATIVE
PHOSPHORYLATION
SITES
WITHIN
THE
C-TERMINUS
OF
SYP-4
AND
CHARACTERISATION
OF
MEIOTIC
DEFECTS
IN
SYP-4
PHOSPHOMUTANTS
.
90
5.3
IDENTIFICATION
OF
A
FUNCTIONAL
DOMAIN
WITHIN
THE
C-TERMINUS
OF
SYP-4
IN
VOLVED
IN
CROSSOVER
REGULATION
.
98
5.4
UNDERSTANDING
HOW
THE
C-TERMINUS
OF
SYP-4
REGULATES
CROSSOVER
FORMATION
111
5.5
DISCUSSION
.
124
CONCLUDING
REMARKS
129
APPENDIX
155
APPENDIX
A
-
LIST
OF
C.
ELEGANS
STRAINS
.
157
APPENDIX
B
-
LIST
OF
CRRNA,
REPAIR
TEMPLATE
AND
GENOTYPING
OLIGO
SEQUENCES
.
159
APPENDIX
C
-
EMBRYONIC
LETHALITY
AND
MALE
PROGENY
FOR
ALL
STRAINS
GENERATED.
.
169
APPENDIX
D
-
ECLIP-SEQ:
COLD
MEMBRANES
IMMUNOBLOTTING
.
175
APPENDIX
E
-
LIST
OF
PRIMERS
AND
ADAPTER
OLIGOS
USED
FOR
CDNA
LIBRARY
PREPARATION
177
APPENDIX
F
-
ECLIP-SEQ
SEQUENCING
DATA PROCESSING
PIPELINE
.
179
APPENDIX
G
-
LIST
OF
IDENTIFIED
RNAS
BY
ECLIP-SEQ
.
185
APPENDIX
H
-
LIST
OF
CAENORHABITIS
SPECIES
USED
FOR
SYP-4
MULTIPLE
SEQUENCE
ALIGNMENT
.
191
APPENDIX
I
-
LIST
OF
PROTEINS
IDENTIFIED
BY
MASS
SPECTROMETRY
AFTER
SYP-4:
:HA
IMMUNOPRECIPITATION
.
193
APPENDIX
J
-
YEAST TWO-HYBRID
ASSAY:
YEAST
STRAINS,
PRIMERS
AND
CONSTRUCTS.
.
.
213
APPENDIX
K
-
PCR
STRATEGY
TO
CONFIRM
CROSSES
FOR
MAPPING
RECOMBINATION
SITES
217
LIST
OF
PUBLICATIONS
219 |
| any_adam_object | 1 |
| author | Neves, Ana Rita Rodrigues 1994- |
| author_GND | (DE-588)1310873097 (DE-588)1024182436 |
| author_facet | Neves, Ana Rita Rodrigues 1994- |
| author_role | aut |
| author_sort | Neves, Ana Rita Rodrigues 1994- |
| author_variant | a r r n arr arrn |
| building | Verbundindex |
| bvnumber | BV050036148 |
| ctrlnum | (OCoLC)1410451018 (DE-599)KXP1870994280 |
| format | Thesis Book |
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| id | DE-604.BV050036148 |
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| indexdate | 2025-01-28T17:18:17Z |
| institution | BVB |
| institution_GND | (DE-588)2024349-2 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-035374000 |
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| owner_facet | DE-355 DE-BY-UBR |
| physical | xxii, 219 Seiten Illustrationen, Diagramme 30 cm |
| publishDate | 2023 |
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| publishDateSort | 2023 |
| record_format | marc |
| spelling | Neves, Ana Rita Rodrigues 1994- Verfasser (DE-588)1310873097 aut The synaptonemal complex and its role in crossover regulation during C. elegans meiosis presented by M.Sc. Ana Rita Rodrigues Neves ; referees: Dr. Sara Cuylen-Häring, Prof. Dr. Michael Knop Heidelberg [2023] xxii, 219 Seiten Illustrationen, Diagramme 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Ruprecht-Karls-University Heidelberg 2023 Text Englisch, Zusammenfassung in englischer und deutscher Sprache (DE-588)4113937-9 Hochschulschrift gnd-content Cuylen, Sara (DE-588)1024182436 dgs Universität Heidelberg (DE-588)2024349-2 dgg B:DE-101 application/pdf https://d-nb.info/1311459715/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=035374000&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p dnb 20231212 DE-101 https://d-nb.info/provenance/plan#dnb |
| spellingShingle | Neves, Ana Rita Rodrigues 1994- The synaptonemal complex and its role in crossover regulation during C. elegans meiosis |
| subject_GND | (DE-588)4113937-9 |
| title | The synaptonemal complex and its role in crossover regulation during C. elegans meiosis |
| title_auth | The synaptonemal complex and its role in crossover regulation during C. elegans meiosis |
| title_exact_search | The synaptonemal complex and its role in crossover regulation during C. elegans meiosis |
| title_full | The synaptonemal complex and its role in crossover regulation during C. elegans meiosis presented by M.Sc. Ana Rita Rodrigues Neves ; referees: Dr. Sara Cuylen-Häring, Prof. Dr. Michael Knop |
| title_fullStr | The synaptonemal complex and its role in crossover regulation during C. elegans meiosis presented by M.Sc. Ana Rita Rodrigues Neves ; referees: Dr. Sara Cuylen-Häring, Prof. Dr. Michael Knop |
| title_full_unstemmed | The synaptonemal complex and its role in crossover regulation during C. elegans meiosis presented by M.Sc. Ana Rita Rodrigues Neves ; referees: Dr. Sara Cuylen-Häring, Prof. Dr. Michael Knop |
| title_short | The synaptonemal complex and its role in crossover regulation during C. elegans meiosis |
| title_sort | the synaptonemal complex and its role in crossover regulation during c elegans meiosis |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1311459715/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=035374000&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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