Characterizing components of the Dictyostelium centrosome:
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| Main Author: | |
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| Format: | Book |
| Language: | English |
| Published: |
2001
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | München, Univ., Fak. für Biologie, Diss., 2002 |
| Physical Description: | 91 Bl. Ill., graph. Darst. : 21 cm |
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MARC
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| 100 | 1 | |a Zoglmeier, Christine |d 1972- |e Verfasser |0 (DE-588)124319912 |4 aut | |
| 245 | 1 | 0 | |a Characterizing components of the Dictyostelium centrosome |c vorgelegt von Christine Zoglmeier, geb. Daunderer |
| 264 | 1 | |c 2001 | |
| 300 | |a 91 Bl. |b Ill., graph. Darst. : 21 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a München, Univ., Fak. für Biologie, Diss., 2002 | ||
| 650 | 0 | 7 | |a Proteine |0 (DE-588)4076388-2 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Centrosom |0 (DE-588)4295334-0 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Dictyostelium discoideum |0 (DE-588)4012135-5 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Dictyostelium discoideum |0 (DE-588)4012135-5 |D s |
| 689 | 0 | 1 | |a Centrosom |0 (DE-588)4295334-0 |D s |
| 689 | 0 | 2 | |a Proteine |0 (DE-588)4076388-2 |D s |
| 689 | 0 | |5 DE-604 | |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010225928&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-010225928 | |
Record in the Search Index
| _version_ | 1807504154271154176 |
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| adam_text |
CONTENTS
CONTENTS
SUMMARY
4
ZUSAMMENFASSUNG
5
ABBREVIATIONS
7
1.
INTRODUCTION
9
1.1
CENTROSOMES
IN
DIFFERENT
ORGANISMS
9
12
THE
ESSENTIAL
CENTROSOMAL
PROTEIN
Y-TUBULIN
AND
ITS
CYTOSOLIC
BINDING
PARTNERS
11
1.3
THE
CENTROSOME-ASSOCIATED
EF-HAND
PROTEIN
CENTRIN
13
2.
MATERIALS
AND
METHODS
15
2.1
MATERIALS
15
2.1.1
REAGENTS
15
2.1.2
ANTIBODIES
15
2.1.3
ENZYMES
15
2.1.4
ANTIBIOTICS
15
2.1.5
MEDIA
16
2.1.5.1
MEDIA
FOR
CULTIVATION
OF
D.
DISCOIDEUM
16
2.1.5.2
MEDIA
FOR
CULTIVATION
OF
E.
COLI
16
2.1.6
BUFFERS
AND
SOLUTIONS
17
2.1.7
VECTORS
18
2.1.8
BACTERIAL
AND
D.
DISCOIDEUM
STRAINS
18
2.1.9
COMPUTER
PROGRAMMES
19
2.1.10
OTHER
MATERIALS
19
22
MOLECULAR
BIOLOGY
METHODS
19
2.2.1
AGAROSE
GEL
ELECTROPHORESIS
19
2.2.2
DNA
EXTRACTION
FROM
AGAROSE
GELS
19
2
2.3
DETERMINATION
OF
DNA
AND
RNA
CONCENTRATION
19
2.2.4
PREPARATION
OF
PLASMID
DNA
20
2.2.5
POLYMERASE
CHAIN
REACTION
(PCR)
20
2.2.6
REVERSE
TRANSCRIPTION
-PCR
(RT-PCR)
20
2.2.7
OLIGONUCLEOTIDES
21
2.2.8
DNA
CLEAVAGE
WITH
RESTRICTION
ENZYMES
21
2.2.9
PHOSPHATASE
TREATMENT
22
2.2.10
BLUNTING
OF
STICKY
ENDS
WITH
T4
DNA
POLYMERASE
22
2.2.11
LIGATION
OFDNA
INTO
PLASMID
VECTORS
22
2.2.12
PREPARATION
OF
ELECTROCOMPETENTE.
COLI
CELLS
22
CONTENTS
2
2.2.13
ELECTROTRANSFORMATION
OFF.
COLI
CELLS
11.
14
IDENTIFICATION
OF
TRANSFORMED
CLONES
IN
E.
COLI
2
J.
15
PREPARATION
OF
CHROMOSOMAL
DNA
FROM/).
DISCOIDEUM
2.2.16
TRANSFORMATION
AND
CLONING
OF
D.
DISCOIDEUM
22.17
ISOLATION
OF
POLYADENYLATED
RNA
FROM
D.
DISCOIDEUM
2.2.18
ELECTROPHORESIS
OF
RNA
AND
NORTHERN
BLOTTING
2.2.19
SOUTHERN
BLOTTING
23.20
RADIOACTIVE
LABELLING
OF
DNA
PROBES,
HYBRIDIZATION
AND
DETECTION
2.2.21
DIGOXYGENIN
LABELLING
OF
DNA
PROBES,
HYBRIDIZATION
AND
DETECTION
2222
CONSTRUCTION
OF
A
SUBGENOMIC
LIBRARY
ENRICHED
M
DDSPC98
SEQUENCE
23.23
SCREENING
OF
CDNA
AND
GENOMIC
LIBRARIES
2.2.24
CONSTRUCTION
OF
THE
VECTORS
FOR
MYC/6XHIS
-TAGGING
OF
ENDOGENOUS
Y-TUBULIN
AND
DDCRP
12.25
CONSTRUCTION
OF
BACTERIAL
EXPRESSION
VECTORS
2.3.
BIOCHEMICAL
AND
IMMUNOLOGICAL
METHODS
2.3.1
SDS-POIYACRYLAMIDE
GEL
ELECTROPHORESIS
(PAGE)
2.3.2
COOMASSIE
STAINING
2.3.3
COLLOIDAL
COOMASSIE
STAINING
2.3.4 METHANOL/CHLOROFORM
PRECIPITATION
OF
PROTEINS
2.3.5
WESTERN
BLOTS
AND
IMMUNOSTAINING
2.3.6
DETERMINATION
OF
PROTEIN
CONCENTRATION
23.7
PURIFICATION
OF
BACTERIALLY
EXPRESSED
PROTEINS
2.3.7.1
PURIFICATION
OF
BACTENALLY
EXPRESSED
HIS
-DDCRP
23.7.2
PURIFICATION
OF
BACTENALLY
EXPRESSED,
MBP-TAGGED
PROTEINS
23.7.3
PURIFICATION
OF
MYC/6XHIS
-TAGGED
CYTOSOLIC
"/-TUBULIN
COMPLEXES
FROM
D.
DISCOIDEUM
2.3.8
PREPARATION
OF
WHOLE
CELL
EXTRACTS,
NUCLEI
AND
CENTROSOMES
FROM
D.
DISCOIDEUM
23.9
GEL
FILTRATION
2.3.10
ANTIGEN
PREPARATION
AND
IMMUNIZATIONS
2.3.11
COVALENT
COUPLING
OF
ANTIBODIES
AND
PURIFIED
PROTEINS
TO
NHS-ACTIVATED
SEPHAROSE
23.12
AFFINITY
PURIFICATION
OF
ANTISERA
2.3.13
IMMUNOPRECIPITATION
2.4
CELL
BIOLOGICAL
METHODS
35
2.4.1
CULTIVATION
AND
PRESERVATION
OF
D.
DISCOIDEUM
35
2.4.2
CULTIVATION
AND
PRESERVATION
OF
E.
COLI
35
2.43
INDIRECT
IMMUNOFLUORESCENCE
MICROSCOPY
ON
WHOLE
D.
DISCOIDEUM
CELLS
AND
ISOLATED
CENTROSOMES
35
2.4.4
CONFOCAL
MICROSCOPY
36
2.4.5
ELECTRON
MICROSCOPY
36
CONTENTS
3
3.
RESULTS
37
3.1
INVESTIGATION
OF
CYTOSOLIC
Y-TUBULIN
COMPLEXES
IN
D.
DISCOIDEUM
37
3.1.1
CLONING
OF
DDSPC97
AND
DDSPC98
37
3.1.2
SEQUENCE
ANALYSIS
OF
DDSPC97
AND
DDSPC98
42
3.1.3
SOUTHERN
AND
NORTHERN
BLOT
ANALYSIS
OF
DDSPC98
AND
DDSPC97
45
3.1.4
INVESTIGATION
OF
THE
SUBCELLULAR
DISTRIBUTION
OF
DDSPC97
AND
DDSPC98
46
3.1.4.1
GENERATION
OF
POLYCLONAL
ANTIBODIES
AND
GFP-TAGGED
MUTANTS
46
3.1.4.2
WESTERN
BLOTS,
IMMUNOFLUORESCENCE
MICROSCOPY
AND
ELECTRON
MICROSCOPY
47
3.1.5
GENERATION
OF
A
MUTANT
STABLY
EXPRESSING
MYC/6XHIS-TAGGED
Y-TUBULIN
52
3.1.6
GEL
FILTRATION
OF
PURIFIED
Y-TUBULIN
COMPLEXES
54
3.1.7
PURIFICATION
AND
IDENTIFICATION
OF
THE
COMPONENTS
OF
D.
DISCOIDEUMY-X^NLIN
COMPLEXES
56
3.2
INVESTIGATION
OF
THE
D.
DISCOIDEUM
HOMOLOGUE
OF
CENTRIN
59
32.
1
CLONING
OF
DDCRP
59
3
21
SOUTHERN
AND
NORTHERN
BLOT
ANALYSIS
OF
DDCRP
60
3.2.3
SEQUENCE
ANALYSIS
OF
DDCRP
61
3.2.4
INVESTIGATION
OF
THE
SUBCELLULAR
DISTRIBUTION
OF
DDCRP
64
3.2.4.1
GENERATION
OF
POLYCLONAL
ANTIBODIES
RAISED
AGAINST
DDCRP
64
3.14.2
SUBCELLULAR
DISTRIBUTION
OF
DDCRP
ON
WESTERN
BLOTS
AND
IN
IMMUNOFLUORESCENCE
MICROSCOPY
65
32.4.3
GENERATION
OF
A
D.
DISCOIDEUM
MUTANT
EXPRESSING
MYC/6XHIS
TAGGED
DDCIP
68
32.4.4
CELL
CYCLE
DEPENDENT
CHANGES
IN
THE
LOCALIZATION
OF
DDCRP
69
4.
DISCUSSION
72
4.1
INVESTIGATION
OF
CYTOSOLIC
Y-TUBULIN
COMPLEXES
IN
D.
DISCOIDEUM
72
4.1.1
TAGGING
OF
ENDOGENOUS
Y-TUBULIN
WITH
A
MYC/6XHIS
TAG
72
4.12
THE
SIZE
AND
STABILITY
OF
D
DISCOIDEUM
Y-TUBULIN
COMPLEXES
72
4.1.3
THE
COMPOSITION
OF
THE
PREVAILING!),
DISCOIDEUM
Y-TUBULIN
COMPLEX
74
4.1.4
THE
POSSIBLE
SIGNIFICANCE
OF
Y-TUBULIN
COMPLEXES
IND.
DISCOIDEUM
75
42
INVESTIGATION
OF
THE
D.
DISCOIDEUM
HOMOLOGUE
OF
CENTRIN
78
42.1
DDCRP
AS
THE
MOST
DIVERGENT
MEMBER
OF
THE
CENTRIN
FAMILY
78
4.2
2
DDCRP
LACKS
TWO
OF
THE
FOUR
EF-HAND
CONSENSUS
MOTIFS
79
42.3
CELL-CYCLE
DEPENDENT
CHANGES
OF
DDCRP
LOCALIZATION
79
42.4
POSSIBLE
INPLICATIONS
FOR
DDCRP
81
5.
CONCLUSION
82
PREFERENCES
83
LEBENSLAUF
91
DANKSAGUNGEN
92 |
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| author_GND | (DE-588)124319912 |
| author_facet | Zoglmeier, Christine 1972- |
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| bvnumber | BV016553482 |
| ctrlnum | (OCoLC)53419050 (DE-599)BVBBV016553482 |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV016553482 |
| illustrated | Illustrated |
| indexdate | 2024-08-16T01:04:48Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-010225928 |
| oclc_num | 53419050 |
| open_access_boolean | |
| owner | DE-19 DE-BY-UBM DE-12 |
| owner_facet | DE-19 DE-BY-UBM DE-12 |
| physical | 91 Bl. Ill., graph. Darst. : 21 cm |
| publishDate | 2001 |
| publishDateSearch | 2001 |
| publishDateSort | 2001 |
| record_format | marc |
| spelling | Zoglmeier, Christine 1972- Verfasser (DE-588)124319912 aut Characterizing components of the Dictyostelium centrosome vorgelegt von Christine Zoglmeier, geb. Daunderer 2001 91 Bl. Ill., graph. Darst. : 21 cm txt rdacontent n rdamedia nc rdacarrier München, Univ., Fak. für Biologie, Diss., 2002 Proteine (DE-588)4076388-2 gnd rswk-swf Centrosom (DE-588)4295334-0 gnd rswk-swf Dictyostelium discoideum (DE-588)4012135-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Dictyostelium discoideum (DE-588)4012135-5 s Centrosom (DE-588)4295334-0 s Proteine (DE-588)4076388-2 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010225928&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Zoglmeier, Christine 1972- Characterizing components of the Dictyostelium centrosome Proteine (DE-588)4076388-2 gnd Centrosom (DE-588)4295334-0 gnd Dictyostelium discoideum (DE-588)4012135-5 gnd |
| subject_GND | (DE-588)4076388-2 (DE-588)4295334-0 (DE-588)4012135-5 (DE-588)4113937-9 |
| title | Characterizing components of the Dictyostelium centrosome |
| title_auth | Characterizing components of the Dictyostelium centrosome |
| title_exact_search | Characterizing components of the Dictyostelium centrosome |
| title_full | Characterizing components of the Dictyostelium centrosome vorgelegt von Christine Zoglmeier, geb. Daunderer |
| title_fullStr | Characterizing components of the Dictyostelium centrosome vorgelegt von Christine Zoglmeier, geb. Daunderer |
| title_full_unstemmed | Characterizing components of the Dictyostelium centrosome vorgelegt von Christine Zoglmeier, geb. Daunderer |
| title_short | Characterizing components of the Dictyostelium centrosome |
| title_sort | characterizing components of the dictyostelium centrosome |
| topic | Proteine (DE-588)4076388-2 gnd Centrosom (DE-588)4295334-0 gnd Dictyostelium discoideum (DE-588)4012135-5 gnd |
| topic_facet | Proteine Centrosom Dictyostelium discoideum Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010225928&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT zoglmeierchristine characterizingcomponentsofthedictyosteliumcentrosome |