Investigation of the polyamine metabolism of Caenorhabditis elegans:
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| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
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2020
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| 100 | 1 | |a Kahl, Janina |d 1983- |e Verfasser |0 (DE-588)1216729298 |4 aut | |
| 245 | 1 | 0 | |a Investigation of the polyamine metabolism of Caenorhabditis elegans |c Janina Kahl |
| 264 | 1 | |a Münster |c 2020 | |
| 300 | |a VII, 99 Blätter |b Illustrationen |c 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Westfälische Wilhelms-Universität Münster |d 2020 |g Beschränkt für den Austausch | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
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| 999 | |a oai:aleph.bib-bvb.de:BVB01-032383476 | ||
Datensatz im Suchindex
| _version_ | 1804181905438932992 |
|---|---|
| adam_text | TABLE
OF
CONTENTS
LIST
OF
ABBREVIATIONS
................................................................................................
IV
LIST
OF
FIGURES
..........................................................................................................
VI
LIST
OF
TABLES
..........................................................................................................
VII
1.
INTRODUCTION
...........................................................................................................
1
1.1.
NEMATODES
.....................................................................................................
1
1.2.
CAENORHABDITIS
ELEGANS
-
A
MODEL
ORGANISM
..................................................
1
1.3.
POLYAMINES
.....................................................................................................
4
1.4.
POLYAMINE
METABOLISM
....................................................................................
5
1.5.
REGULATION
OF
POLYAMINE
HOMEOSTASIS
.............................................................
7
1.6.
THE
POLYAMINE
METABOLISM
IN
NEMATODES
........................................................
8
1.7.
POLYAMINES
AND
STRESS
.................................................................................
14
1.8.
AIMS
.............................................................................................................
17
2.
MATERIALS
&
METHODS
...........................................................................................
18
2.1.
MATERIALS
.......................................................................................................
18
2.1.1.
LABORATORY
EQUIPMENT
...............................................................................
18
2.1.2.
CHEMICALS
.................................................................................................
19
2.1.3.
ORGANISMS
.................................................................................................
20
2.1.4.
OLIGONUCLEOTIDES........................................................................................
21
2.1.5.
KIT
SYSTEMS
................................................................................................
22
2.1.6.
ENZYMES
...................................................................................................
22
2.1.7.
BUFFER,
MEDIA
AND
SOLUTIONS
......................................................................
23
2.1.8.
SOFTWARE
....................................................................................................
24
2.2.
METHODS
.........................................................................................................
25
2.2.1.
GENERAL
PROCEDURES...................................................................................
25
2.2.1.1.
CULTIVATION
OF
E.
COLI
...............................................................................
25
2.2.1.2.
CULTIVATION
AND
MAINTENANCE
OF
C.
ELEGANS
............................................
25
T
ABLE
OF
CONTENTS
2.2.1.3.
SINGLE
WORM
LYSIS
....................................................................................
25
2.2.1.4.
PCR
........................................................................................................
25
2.2.1.5.
RNA
ISOLATION
..........................................................................................
26
2.2.1.6.
CDNA
SYNTHESIS
.......................................................................................
26
2.2.1.7.
SEQUENCING
.............................................................................................
26
2.2.1.8.
STATISTICS..................................................................................................
26
2.2.2.
GENERATING
OAZ-1
MUTANTS
VIA
CRISPR/CAS9
...........................................
26
2.2.3.
DETERMINATION
OF
C.
ELEGANS
LIFE
TRAIT
PARAMETERS
......................................
30
2.2.4.
ANALYSIS
OF
THE
C.
ELEGANS
OAZ-1,
ODC-1
AND
AZIN-1
EXPRESSION
PATTERN...
31
2.2.5.
DEVELOPMENT
OF
THE
SPECIFIC
REPORTER
P
OAZ-1
::GFP::OAZ-1
::MCHERRY
..........
31
2.2.6.
POLYAMINE
QUANTIFICATION
BY
HIGH
PERFORMANCE
LIQUID
CHROMATOGRAPHY
(HPLC)
............................................................................................................
32
2.2.7.
QUANTITATIVE
REAL-TIME
PCR
FOR
TRANSCRIPTIONAL
ANALYSIS
OF
POLYAMINE
METABOLISM
GENES
..................................................................................................
33
3.
RESULTS
................................................................................................................
34
3.1.
GENERATION
OF
A
FRAMESHIFT-INDEPENDENT
MUTANT
VIA
CRISPR/CAS9
...............
34
3.2.
CRISPR/CAS9
GENERATED
C.
ELEGANS
OAZ-1
DELETION
MUTANTS
ARE
VIABLE
....
36
3.3.
KNOCKOUT
OF
OAZ-1
AFFECTS
BROOD
SIZE,
WHILE
LOSS
OF
THE
REGULATORY
FRAMESHIFT
SITE
RESULTS
IN
A
PROLONGED
LIFESPAN
........................................................................
41
3.4.
KNOCKOUT
OF
AZIN-1
AFFECTS
LIFESPAN,
DEVELOPMENT
AND
REPRODUCTION
RATE
....
43
3.5.
POLYAMINE
METABOLISM
GENES
ARE
MAINLY
EXPRESSED
IN
THE
INTESTINE
..........
44
3.6.
SPATIAL
AND
TEMPORAL
EXPRESSION
PATTERN
OF
KEY
POLYAMINE
METABOLISM
GENES
DURING
DEVELOPMENT................................................................................................
47
3.7.
SPERMIDINE
AND
SPERMINE
INDUCE
TRANSLATIONAL
FRAMESHIFTING
OF
OAZ-1
MRNA.
49
3.8.
COMPARATIVE
TRANSCRIPTIONAL
ANALYSIS
OF
POLYAMINE
METABOLISM
GENES
.......
53
3.9.
STRESS
INDUCES
OAZ-1
EXPRESSION
..................................................................
54
3.10.
OPTIMIZATION
OF
POLYAMINE
QUANTIFICATION
IN
C.
ELEGANS
.............................
56
II
T
ABLE
OF
CONTENTS
3.11.
STRESS
DECREASES
POLYAMINE
CONTENT
IN
C.
ELEGANS
...................................
58
3.12.
DETERMINATION
OF
POLYAMINE
CONTENT
IN
DAPHNIA
MAGNA
...........................
60
4.
DISCUSSION...........................................................................................................
62
4.1.
THE
REGULATORY
NETWORK
OF
POLYAMINE
METABOLISM
IN
C.
ELEGANS
.................
62
4.1.1.
GENERATION
OF
AN
OAZ-1
KNOCKOUT
MUTANT
VIA
CRISPR/CAS9
....................
63
4.1.2.
THE
EFFECT
OF
DISRUPTING
THE
REGULATORY
NETWORK
OF
POLYAMINE
METABOLISM
ON
LIFETRAIT
PARAMETERS
.................................................................................................
63
4.1.3.
GENE
EXPRESSION
OF
POLYAMINE
METABOLISM
GENES
....................................
66
4.1.4.
THE
EFFECT
OF
POLYAMINES
ON
OAZ-1
AND
ODC-1
EXPRESSION
........................
68
4.2.
OPTIMIZATION
AND
SUBSEQUENT
POLYAMINE
ANALYSIS
IN
C.
ELEGANS
.................
70
4.3.
POLYAMINE
ANALYSIS
IN
THE
MODEL
ORGANISM
D.
MAGNA
..................................
71
4.4.
THE
ROLE
OF
POLYAMINE
METABOLISM
IN
STRESS
RESPONSE
..................................
72
5.
SUMMARY
.............................................................................................................
75
6.
OUTLOOK
................................................................................................................
77
REFERENCES
................................................................................................................
79
APPENDIX
....................................................................................................................
94
SEQUENCE
ALIGNMENTS.............................................................................................
94
VECTOR
MAPS
...........................................................................................................
95
DANKSAGUNG
...........................................................................................................
96
CURRICULUM
VITAE
.....................................................................................................
98
III
|
| adam_txt |
TABLE
OF
CONTENTS
LIST
OF
ABBREVIATIONS
.
IV
LIST
OF
FIGURES
.
VI
LIST
OF
TABLES
.
VII
1.
INTRODUCTION
.
1
1.1.
NEMATODES
.
1
1.2.
CAENORHABDITIS
ELEGANS
-
A
MODEL
ORGANISM
.
1
1.3.
POLYAMINES
.
4
1.4.
POLYAMINE
METABOLISM
.
5
1.5.
REGULATION
OF
POLYAMINE
HOMEOSTASIS
.
7
1.6.
THE
POLYAMINE
METABOLISM
IN
NEMATODES
.
8
1.7.
POLYAMINES
AND
STRESS
.
14
1.8.
AIMS
.
17
2.
MATERIALS
&
METHODS
.
18
2.1.
MATERIALS
.
18
2.1.1.
LABORATORY
EQUIPMENT
.
18
2.1.2.
CHEMICALS
.
19
2.1.3.
ORGANISMS
.
20
2.1.4.
OLIGONUCLEOTIDES.
21
2.1.5.
KIT
SYSTEMS
.
22
2.1.6.
ENZYMES
.
22
2.1.7.
BUFFER,
MEDIA
AND
SOLUTIONS
.
23
2.1.8.
SOFTWARE
.
24
2.2.
METHODS
.
25
2.2.1.
GENERAL
PROCEDURES.
25
2.2.1.1.
CULTIVATION
OF
E.
COLI
.
25
2.2.1.2.
CULTIVATION
AND
MAINTENANCE
OF
C.
ELEGANS
.
25
T
ABLE
OF
CONTENTS
2.2.1.3.
SINGLE
WORM
LYSIS
.
25
2.2.1.4.
PCR
.
25
2.2.1.5.
RNA
ISOLATION
.
26
2.2.1.6.
CDNA
SYNTHESIS
.
26
2.2.1.7.
SEQUENCING
.
26
2.2.1.8.
STATISTICS.
26
2.2.2.
GENERATING
OAZ-1
MUTANTS
VIA
CRISPR/CAS9
.
26
2.2.3.
DETERMINATION
OF
C.
ELEGANS
LIFE
TRAIT
PARAMETERS
.
30
2.2.4.
ANALYSIS
OF
THE
C.
ELEGANS
OAZ-1,
ODC-1
AND
AZIN-1
EXPRESSION
PATTERN.
31
2.2.5.
DEVELOPMENT
OF
THE
SPECIFIC
REPORTER
P
OAZ-1
::GFP::OAZ-1
::MCHERRY
.
31
2.2.6.
POLYAMINE
QUANTIFICATION
BY
HIGH
PERFORMANCE
LIQUID
CHROMATOGRAPHY
(HPLC)
.
32
2.2.7.
QUANTITATIVE
REAL-TIME
PCR
FOR
TRANSCRIPTIONAL
ANALYSIS
OF
POLYAMINE
METABOLISM
GENES
.
33
3.
RESULTS
.
34
3.1.
GENERATION
OF
A
FRAMESHIFT-INDEPENDENT
MUTANT
VIA
CRISPR/CAS9
.
34
3.2.
CRISPR/CAS9
GENERATED
C.
ELEGANS
OAZ-1
DELETION
MUTANTS
ARE
VIABLE
.
36
3.3.
KNOCKOUT
OF
OAZ-1
AFFECTS
BROOD
SIZE,
WHILE
LOSS
OF
THE
REGULATORY
FRAMESHIFT
SITE
RESULTS
IN
A
PROLONGED
LIFESPAN
.
41
3.4.
KNOCKOUT
OF
AZIN-1
AFFECTS
LIFESPAN,
DEVELOPMENT
AND
REPRODUCTION
RATE
.
43
3.5.
POLYAMINE
METABOLISM
GENES
ARE
MAINLY
EXPRESSED
IN
THE
INTESTINE
.
44
3.6.
SPATIAL
AND
TEMPORAL
EXPRESSION
PATTERN
OF
KEY
POLYAMINE
METABOLISM
GENES
DURING
DEVELOPMENT.
47
3.7.
SPERMIDINE
AND
SPERMINE
INDUCE
TRANSLATIONAL
FRAMESHIFTING
OF
OAZ-1
MRNA.
49
3.8.
COMPARATIVE
TRANSCRIPTIONAL
ANALYSIS
OF
POLYAMINE
METABOLISM
GENES
.
53
3.9.
STRESS
INDUCES
OAZ-1
EXPRESSION
.
54
3.10.
OPTIMIZATION
OF
POLYAMINE
QUANTIFICATION
IN
C.
ELEGANS
.
56
II
T
ABLE
OF
CONTENTS
3.11.
STRESS
DECREASES
POLYAMINE
CONTENT
IN
C.
ELEGANS
.
58
3.12.
DETERMINATION
OF
POLYAMINE
CONTENT
IN
DAPHNIA
MAGNA
.
60
4.
DISCUSSION.
62
4.1.
THE
REGULATORY
NETWORK
OF
POLYAMINE
METABOLISM
IN
C.
ELEGANS
.
62
4.1.1.
GENERATION
OF
AN
OAZ-1
KNOCKOUT
MUTANT
VIA
CRISPR/CAS9
.
63
4.1.2.
THE
EFFECT
OF
DISRUPTING
THE
REGULATORY
NETWORK
OF
POLYAMINE
METABOLISM
ON
LIFETRAIT
PARAMETERS
.
63
4.1.3.
GENE
EXPRESSION
OF
POLYAMINE
METABOLISM
GENES
.
66
4.1.4.
THE
EFFECT
OF
POLYAMINES
ON
OAZ-1
AND
ODC-1
EXPRESSION
.
68
4.2.
OPTIMIZATION
AND
SUBSEQUENT
POLYAMINE
ANALYSIS
IN
C.
ELEGANS
.
70
4.3.
POLYAMINE
ANALYSIS
IN
THE
MODEL
ORGANISM
D.
MAGNA
.
71
4.4.
THE
ROLE
OF
POLYAMINE
METABOLISM
IN
STRESS
RESPONSE
.
72
5.
SUMMARY
.
75
6.
OUTLOOK
.
77
REFERENCES
.
79
APPENDIX
.
94
SEQUENCE
ALIGNMENTS.
94
VECTOR
MAPS
.
95
DANKSAGUNG
.
96
CURRICULUM
VITAE
.
98
III |
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| any_adam_object_boolean | 1 |
| author | Kahl, Janina 1983- |
| author_GND | (DE-588)1216729298 |
| author_facet | Kahl, Janina 1983- |
| author_role | aut |
| author_sort | Kahl, Janina 1983- |
| author_variant | j k jk |
| building | Verbundindex |
| bvnumber | BV046975338 |
| ctrlnum | (OCoLC)1195771843 (DE-599)DNB1216068380 |
| discipline | Biologie |
| discipline_str_mv | Biologie |
| format | Thesis Book |
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| id | DE-604.BV046975338 |
| illustrated | Illustrated |
| index_date | 2024-07-03T15:48:25Z |
| indexdate | 2024-07-10T08:59:05Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-032383476 |
| oclc_num | 1195771843 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | VII, 99 Blätter Illustrationen 30 cm |
| publishDate | 2020 |
| publishDateSearch | 2020 |
| publishDateSort | 2020 |
| record_format | marc |
| spelling | Kahl, Janina 1983- Verfasser (DE-588)1216729298 aut Investigation of the polyamine metabolism of Caenorhabditis elegans Janina Kahl Münster 2020 VII, 99 Blätter Illustrationen 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Westfälische Wilhelms-Universität Münster 2020 Beschränkt für den Austausch (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1216068380/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032383476&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Kahl, Janina 1983- Investigation of the polyamine metabolism of Caenorhabditis elegans |
| subject_GND | (DE-588)4113937-9 |
| title | Investigation of the polyamine metabolism of Caenorhabditis elegans |
| title_auth | Investigation of the polyamine metabolism of Caenorhabditis elegans |
| title_exact_search | Investigation of the polyamine metabolism of Caenorhabditis elegans |
| title_exact_search_txtP | Investigation of the polyamine metabolism of Caenorhabditis elegans |
| title_full | Investigation of the polyamine metabolism of Caenorhabditis elegans Janina Kahl |
| title_fullStr | Investigation of the polyamine metabolism of Caenorhabditis elegans Janina Kahl |
| title_full_unstemmed | Investigation of the polyamine metabolism of Caenorhabditis elegans Janina Kahl |
| title_short | Investigation of the polyamine metabolism of Caenorhabditis elegans |
| title_sort | investigation of the polyamine metabolism of caenorhabditis elegans |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1216068380/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032383476&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT kahljanina investigationofthepolyaminemetabolismofcaenorhabditiselegans |
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