The absence of nidogen-1 dous not affect murine basement membrane formation:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
2000
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Köln, Univ., Diss., 2001 |
| Beschreibung: | 90 S. Ill., graph. Darst. : 21 cm |
Internformat
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| 245 | 1 | 0 | |a The absence of nidogen-1 dous not affect murine basement membrane formation |c vorgelegt von Monzur Murshed |
| 264 | 1 | |c 2000 | |
| 300 | |a 90 S. |b Ill., graph. Darst. : 21 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Köln, Univ., Diss., 2001 | ||
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| 650 | 0 | 7 | |a Entactin |0 (DE-588)4316135-2 |2 gnd |9 rswk-swf |
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| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-009406830 | |
Datensatz im Suchindex
| _version_ | 1807593549467746304 |
|---|---|
| adam_text |
INDEX
1.
INTRODUCTION
1
1.1
THE
BASEMENT
MEMBRANE
1
1.2
BASEMENT
MEMBRANE
COMPONENTS
1
1.3
NIDOGENS
4
1.4
LAMININ-NIDOGEN
COMPLEX
7
1.5
BASEMENT
MEMBRANE
ASSEMBLY
8
1.6
MUTATIONS
IN
BASEMENT
MEMBRANE
COMPONENTS
11
1.7
AIMS
OF
THE
STUDY
13
2.
MATERIALS
AND
METHODS
15
2.1
MATERIALS
15
2.1.1
CHEMICALS
15
2.1.2
BUFFERS
15
2.1.3
BACTERIAL
CULTURE
17
2.1.3.1
MEDIA
17
2.1.3.2
BACTERIAL
STRAIN
17
2.1.4
PLASMIDS
17
2.1.5
EMBRYONIC
STEM
CELL
CULTURE
18
2.1.5.1
MEDIA
18
2.1.5.2
EMBRYONIC
STEM
CELLS
AND
EMBRYONIC
FIBROBLASTS
19
2.1.6
SYNTHETIC
OLIGONUCLEOTIDES
19
2.1.7
PRIMARY
AND
SECONDARY
ANTIBODIES
20
2.2
METHODS
20
2.2.1
RECOMBINANT
DNA
TECHNIQUES
20
2.2.1.1
RESTRICTION
ENZYME
DIGESTION,
DNA
PURIFICATION,
AND
LIGATION
20
2.2.2
GENE
TARGETING
IN
EMBRYONIC
STEM
CELLS
21
2.2.2.1
MYTOMYCIN
C
TREATMENT
OF
EMBRYONIC
FIBROBLASTS
21
2.2.2.2
EMBRYONIC
STEM
CELL
CULTURE
21
2.2.2.3
EMBRYONIC
STEM
CELL
TRANSFECTION
22
2.2.2.4
ANTIBIOTIC
SELECTION
AND
PICKING
OF
TRANSFECTED
CELLS
22
2.2.2.5
SPLITTING
AND
FREEZING
OF
EMBRYONIC
STEM
CELLS
23
2.2.2.6
CRE
RECOMBINASE
MEDIATED
DELETION
23
2.2.3
NUCLEIC
ACID
ANALYSIS
24
2.2.3.1
ISOLATION
OF
EMBRYONIC
STEM
CELL
DNA
24
2.2.3.2
ISOLATION
OF
GENOMIC
DNA
FROM
MOUSE
TAILS
24
2.2.3.3
SOUTHERN
BLOT
ANALYSIS
25
2.2.3.4
POLYMERASE
CHAIN
REACTION
(PCR)
25
2.2.3.5
RNA
ISOLATION
FROM
MOUSE
TISSUES
26
2.2.3.6
NORTHERN
BLOT
ANALYSIS
26
2.2.4
PROTEIN
ANALYSIS
METHODS
27
2.2.4.1
PROTEIN
EXTRACTION
FROM
MOUSE
TISSUES
27
2.2.4.2
EXTRACTION
OF
LAMININ
FROM
MOUSE
HEART
27
2.2.4.3
SDS-POLYACRYLAMIDE
GEL
ELECTROPHORESIS
AND
WESTERN
BLOT
ANALYSIS
28
2.2.5
GENERATION
OF
CHIMERIC
ANIMALS
AND
HOMOZYGOUS
OFFSPRING
28
2.2.6
GENERATION
OF
EMBRYOID
BODIES
(EBS)
29
2.2.7
PREPARATION
OF
FROZEN
SECTIONS
30
2.2.8
IMMUNOFLUORESCENCE
STAINING
OF
FROZEN
SECTIONS
30
2.2.9
ELECTROM
MICROSCOPY
30
2.2.10 STAINING
OF
WHOLE
MOUNT
EMBRYOS
30
3.
RESULTS
32
3.1
TARGETING
THE
N1D-1
LOCUS
32
3.1.1
ISOLATION
OF
N1D-1
GENOMIC
SEQUENCES
33
3.1.2
CONSTRUCTION
OF
THE
TARGETING
VECTOR
PNIDMUTL
34
3.1.3
CONSTRUCTION
OF
THE
TARGETING
VECTOR
PLAC
I
36
3.1.4
GENERATION
OF
ES
CELL
CLONES
CARRYING
A
MUTATED
N1D-1
LOCUS
38
3.1.4.1
SCREENING
OF
PNIDMUTL
TRANSFECTED
ES
CELL
CLONES
38
3.1.4.2
RECLONING
OFES
CELL
CLONES
T2/64
AND
T2/140
42
3.1.4.3
SCREENING
OF
PLAC
1
TRANSFECTED
ES
CELL
CLONES
42
3.1.5
ANALYSIS
OF
ES
CELL
CLONES
AFTER
CRE
RECOMBINASE
MEDIATED
DELETION
44
3.2
ANALYSIS
OF
NID-1
DISRUPTION
IN
VITRO
46
3.2.1
TARGETED
DISRUPTION
OF
BOTH
NID-1
ALLELES
46
3.2.2
ANALYSIS
OF
EMBRYOID
BODIES
48
3.3
GERM
LINE
TRANSMISSION
OF
THE
TARGETED
ALLELE
51
3.4
GENOTYPING
OF
OFFSPRING
FROM
THE
BREEDING
OF
HETEROZYGOUS
ANIMALS
52
3.5
LACZ
STAINING
OF
WHOLE
MOUNT
EMBRYOS
55
3.6
ANALYSIS
OF
MICE
HOMOZYGOUS
FOR
THE
NID-1
MUTATIONS
55
3.6.1
ANALYSIS
OF
NIDOGEN-1
EXPRESSION
IN-/-ANIMALS
56
3.6.2
IMMUNOFLUORESCENCE
STUDIES
ON
DIFFERENT
-/
TISSUES
57
3.6.3
ANALYSIS
OFNIDOGEN-2
EXPRESSION
IN
NIDOGEN-1
DEFICIENT
ANIMALS
59
3.6.4
IMMUNOBLOT
ANALYSIS
OF
LAMININ
IN
-/
TISSUE
EXTRACTS
61
3.6.5
EXTRACTION
OF
LAMININ
FROM
+/+
AND
-/
MOUSE
HEARTS
62
3.7
BASEMENT
MEMBRANE
FORMATION
IN
THE
ABSENCE
OF
NIDOGEN-L
65
4.
DISCUSSIONS
67
4.1
TARGETED
DISRUPTION
OF
NID-1
GENES
IN
ES
CELL
67
4.2 ANALYSIS
OF
NID-1
DISRUPTION
IN
VITRO
71
4.3
GENERATION
OF
MOUSE
LINES
WITH
DISRUPTED
NID-1
GENES
73
4.4
CONSEQUENCES
OF
NID-1
DISRUPTION
IN
VIVO
75
5.
SUMMARY
80
6.
ZUSAMMENFASSUNG
81
7.
REFERENCES
82 |
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| author | Murshed, Monzur |
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| indexdate | 2024-08-17T00:45:42Z |
| institution | BVB |
| language | English |
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| open_access_boolean | |
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| owner_facet | DE-355 DE-BY-UBR |
| physical | 90 S. Ill., graph. Darst. : 21 cm |
| publishDate | 2000 |
| publishDateSearch | 2000 |
| publishDateSort | 2000 |
| record_format | marc |
| spelling | Murshed, Monzur Verfasser aut The absence of nidogen-1 dous not affect murine basement membrane formation vorgelegt von Monzur Murshed 2000 90 S. Ill., graph. Darst. : 21 cm txt rdacontent n rdamedia nc rdacarrier Köln, Univ., Diss., 2001 Maus - Basalmembran - Blutstammzelle - LacZ - Genmutation - Entactin Maus (DE-588)4169148-9 gnd rswk-swf LacZ (DE-588)4311588-3 gnd rswk-swf Blutstammzelle (DE-588)4146093-5 gnd rswk-swf Entactin (DE-588)4316135-2 gnd rswk-swf Basalmembran (DE-588)4144086-9 gnd rswk-swf Genmutation (DE-588)4156638-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Maus (DE-588)4169148-9 s Basalmembran (DE-588)4144086-9 s Blutstammzelle (DE-588)4146093-5 s LacZ (DE-588)4311588-3 s Genmutation (DE-588)4156638-5 s Entactin (DE-588)4316135-2 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009406830&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Murshed, Monzur The absence of nidogen-1 dous not affect murine basement membrane formation Maus - Basalmembran - Blutstammzelle - LacZ - Genmutation - Entactin Maus (DE-588)4169148-9 gnd LacZ (DE-588)4311588-3 gnd Blutstammzelle (DE-588)4146093-5 gnd Entactin (DE-588)4316135-2 gnd Basalmembran (DE-588)4144086-9 gnd Genmutation (DE-588)4156638-5 gnd |
| subject_GND | (DE-588)4169148-9 (DE-588)4311588-3 (DE-588)4146093-5 (DE-588)4316135-2 (DE-588)4144086-9 (DE-588)4156638-5 (DE-588)4113937-9 |
| title | The absence of nidogen-1 dous not affect murine basement membrane formation |
| title_auth | The absence of nidogen-1 dous not affect murine basement membrane formation |
| title_exact_search | The absence of nidogen-1 dous not affect murine basement membrane formation |
| title_full | The absence of nidogen-1 dous not affect murine basement membrane formation vorgelegt von Monzur Murshed |
| title_fullStr | The absence of nidogen-1 dous not affect murine basement membrane formation vorgelegt von Monzur Murshed |
| title_full_unstemmed | The absence of nidogen-1 dous not affect murine basement membrane formation vorgelegt von Monzur Murshed |
| title_short | The absence of nidogen-1 dous not affect murine basement membrane formation |
| title_sort | the absence of nidogen 1 dous not affect murine basement membrane formation |
| topic | Maus - Basalmembran - Blutstammzelle - LacZ - Genmutation - Entactin Maus (DE-588)4169148-9 gnd LacZ (DE-588)4311588-3 gnd Blutstammzelle (DE-588)4146093-5 gnd Entactin (DE-588)4316135-2 gnd Basalmembran (DE-588)4144086-9 gnd Genmutation (DE-588)4156638-5 gnd |
| topic_facet | Maus - Basalmembran - Blutstammzelle - LacZ - Genmutation - Entactin Maus LacZ Blutstammzelle Entactin Basalmembran Genmutation Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009406830&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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