Characterization of Vav-1 and Vav-2: study of Vav-1 -/- Vav-2 -/- B cells and generation of Vav-1 knock - in mice
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
2002
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Zsfassung in dt. Sprache. - Würzburg, Univ., Diss., 2003 |
| Beschreibung: | 117 Bl. Ill., graph. Darst. |
Internformat
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| 245 | 1 | 0 | |a Characterization of Vav-1 and Vav-2 |b study of Vav-1 -/- Vav-2 -/- B cells and generation of Vav-1 knock - in mice |c von Kerry Tedford |
| 264 | 1 | |c 2002 | |
| 300 | |a 117 Bl. |b Ill., graph. Darst. | ||
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Datensatz im Suchindex
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|---|---|
| adam_text |
TABLE OF CONTENTS
SUMMARY.8
ZUSAMMENFASSUNG.9
INTRODUCTION.II
VAV FAMILY STRUCTURE, EXPRESSION AND ACTIVITY.11
T CELL DEVELOPMENT.16
TCR SIGNALING.17
VAV-1 KNOCKOUT MICE: T CELL DEVELOPMENT PHENOTYPE.20
VAV-1 KNOCKOUT MICE: SIGNALING PHENOTYPE.22
B CELL DEVELOPMENT AND SIGNALING.23
VAV-1 KNOCKOUT MICE: B CELL DEVELOPMENT.25
GOALS OF THE DOCTORAL WORK. 27
RESULTS.27
GENERATION OF VAV-2-MUTANT MICE.27
ANALYSIS OF LYMPHOCYTE POPULATIONS IN VAV-MUTANT MICE.30
DEFECTS IN VAV-MUTANT T AND B CELL ARE INTRINSIC TO LYMPHOCYTES AND
CONFER A
COMPETITIVE DISADVANTAGE IN DEVELOPMENT.34
FUNCTIONAL DEFECTS IN VAV-MUTANT B CELLS.37
IMPAIRED UPREGULATION OF CD86 AND MHC CLASS II IN VAV-1'7' VAV-2'7' B
CELIJS.43
DEGRADATION IN LYSATES.44
VAV-1- AND VAV-2-SH2 DOMAIN-ASSOCIATED PROTEINS IN B CELLS 47
ANALYSIS OF SIGNALING PROTEINS IN VAV-1'
7' VAV-2'7' B CELLS 47
DEFECTIVE CALCIUM MOBILIZATION IN VAV-MUTANT B CELLS 49
AGGREGATION OF BCR AND LIPID RAFTS IN VAV-1'
7' VAV-2'7' B CELLS 50
IN VIVO
MUTATIONAL ANALYSIS OF VAV-I.52
CLONING OF VAV-1 KNOCK/IN TARGETING VECTORS.54
MUTATIONS OF VAV-1 CDNA FOR KNOCK/IN TARGETING VECTORS.56
INTRODUCTION OF VAV-1 KNOCK/IN TARGETING VECTORS INTO MOUSE ES CELLS.58
DISCUSSION.59
VAV-1'
7'VAV-2'7' B CELLS. 59
DEGRADATION IN LYSATES OF SPLENIC CELL POPULATIONS.60
INCREASED IGMHI CELLS IN VAV-1'7' AND VAV-1'7' VAV-2"7 B CELLS.60
DIFFERENCES IN FUNCTIONS REGULATED BY VAV-1 AND VAV-2.61
POTENTIAL ROLES FOR VAV CATALYTIC ACTIVITY IN B CELLS: RHO GTPASES IN B
CELL
CYTOSKELETAL REORGANIZATION AND PIP2 PRODUCTION.62
POTENTIAL ROLES FOR VAV ADAPTOR DOMAINS IN B CELLS: COMPLEX FORMATION.66
GENERATION OF VAV-1 KNOCK/IN MUTANT MICE 67
FUTURE EXPERIMENTAL APPROACHES.
69
METHODS.70
70
MICROBIOLOGICAL METHODS.
BACTERIAL CULTURE
.
70
TRANSFORMATION OF PLASMIDS
.
CLONING.71
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/968427154
PREPARATION OF PLASMID DNA
.
CONCENTRATION OF DNA
.
MEASUREMENT OF DNA CONCENTRATION
.
PHENOL/CHLOROFORM EXTRACTION
.
AGAROSE GEL ELECTROPHORESIS
.
PURIFICATION OF DNA FROM AGAROSE
.
DIGESTION OF DNA
.
DEPHOSPHORYLATION OF DNA WITH CALF INTESTINAL PHOSPHATASE (CIP)
.
FILL-IN OF OVERHANGING DNA ENDS
.
LIGATION OF DNA
.
STANDARD MOLECULAR BIOLOGICAL METHODS.
MUTAGENESIS BY PCR
.
DNA SEQUENCING.
.
GST PROTEIN PREPARATION
.
PROTEIN BIOCHEMISTRY.
LYSIS OF CELLS AND IMMUNOPRECIPITATION
. .
SDS-PAGE, GEL TRANSFER, WESTERN BLOTTING
.
COOMMASSIE
.
CELLULAR BIOLOGICAL METHODS
CENTRIFUGATION OF LYMPHOCYTES
.
COUNTING CELLS
.
CELL PURIFICATION
.
FACS STAINING OF CELLS
.
PROLIFERATION/UPREGULATION OF ACTIVATION MARKERS
.
ANTIGENIC CHALLENGES
.
MEASUREMENT OF INTRACELLULAR CA~ MOBILIZATION
.
CELL STIMULATION, IMMUNOPRECIPITATION AND PROTEIN DETECTION
.
BCR
/
RAFT AGGREGATION
.
EXPRESSION OFVAV-1 AND VAV-2
.
ESCELL WORK.
.
PCR SCREENING OF ES CELL CLONES
.
ANIMAL CARE AND HANDLING
.
EMBRYO STAINING
.
GENERATION OF BONE MARROW CHIMERIC MICE
.
GENERATION OF VAV-2 KNOCKOUT MICE
.
CONSTRUCTION OF THE VAV-1 KNOCK/IN TARGETING VECTORS.
PREPARING PLASMIDS FOR PCR MUTAGENESIS
.
PCR MUTAGENESIS OFVAV-1 CDNA SEGMENTS
.
CONSTRUCTING FULL-LENGTH MUTAGENIZED VAV-1 CDNA WITH OR WITHOUT THE FLAG
TAG
MUTATED VAV-1 CDNA CONSTRUCTS IN EXPRESSION VECTORS
.
PREPARATION OF THE LONG ARM FOR CLONING
.
FUSION OF THE LONG ARM TO THE CDNA AT EXON YYA
.
PREPARATION OF THE TARGETING VECTOR
.
ASSEMBLY OF THE LONG ARM/CDNA FUSION WITH THE TARGETING VECTOR
.
PCR AND SOUTHERN STRATEGIES FOR CHECKING INTEGRATION OF THE KNOCK/IN
TARGETING VECTORS
MATERIALS.
CHEMICALS AND REAGENTS
.
BIOLOGICAL REAGENTS
.
71
71
71
71
72
72
72
72
72
73
73
73
74
74
.74
74
75
75
.76
76
.76
.76
.77
.77
.
7
H
.78
.78
.79
.79
.79
.80
.81
.81
.81
.81
.81
.81
.84
.87
.90
.91
.91
.92
.94
.95
.96
.96
.97
1
PLASMIDS
.
OLIGOMERS
.!.
KITS
.
CONSUMABLE PRODUCTS
.
EQUIPMENT
.
SOFTWARE
.
STATISTICAL ANALYSIS
.
ABBREVIATIONS
.
PRIMERS FOR MUTAGENIZING VAV-L CDNA
.
PRIMERS FOR SEQUENCING VAV-L
.
PRIMER FOR CHECKING VAV-L GENOMIC INTEGRATION-NEO PRIMER
.
PRIMER FOR CHECKING VAV-L GENOMIC INTEGRATION-VAV-L 3
'
EXTERNAL PRIMER
.97
.97
.97
.98
.98
.98
.98
.99
100
101
101
101
APPENDIX 2: TABLE 3 - A LIST OF PLASMIDS USED IN THE CONSTRUCTION OF THE
VAV-1 KNOCK/IN TARGETING VECTORS.1
REFERENCES
107
CURRICULUM VITAE
116
PUBLICATION LIST
117 |
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| spelling | Tedford, Kerry Verfasser aut Characterization of Vav-1 and Vav-2 study of Vav-1 -/- Vav-2 -/- B cells and generation of Vav-1 knock - in mice von Kerry Tedford 2002 117 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. Sprache. - Würzburg, Univ., Diss., 2003 Guanosintriphosphatasen (DE-588)4324625-4 gnd rswk-swf Mikroform (DE-588)4039216-8 gnd rswk-swf Molekularbiologie (DE-588)4039983-7 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Guanosintriphosphatasen (DE-588)4324625-4 s Molekularbiologie (DE-588)4039983-7 s Mikroform (DE-588)4039216-8 s 1\p DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010303062&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk |
| spellingShingle | Tedford, Kerry Characterization of Vav-1 and Vav-2 study of Vav-1 -/- Vav-2 -/- B cells and generation of Vav-1 knock - in mice Guanosintriphosphatasen (DE-588)4324625-4 gnd Mikroform (DE-588)4039216-8 gnd Molekularbiologie (DE-588)4039983-7 gnd |
| subject_GND | (DE-588)4324625-4 (DE-588)4039216-8 (DE-588)4039983-7 (DE-588)4113937-9 |
| title | Characterization of Vav-1 and Vav-2 study of Vav-1 -/- Vav-2 -/- B cells and generation of Vav-1 knock - in mice |
| title_auth | Characterization of Vav-1 and Vav-2 study of Vav-1 -/- Vav-2 -/- B cells and generation of Vav-1 knock - in mice |
| title_exact_search | Characterization of Vav-1 and Vav-2 study of Vav-1 -/- Vav-2 -/- B cells and generation of Vav-1 knock - in mice |
| title_full | Characterization of Vav-1 and Vav-2 study of Vav-1 -/- Vav-2 -/- B cells and generation of Vav-1 knock - in mice von Kerry Tedford |
| title_fullStr | Characterization of Vav-1 and Vav-2 study of Vav-1 -/- Vav-2 -/- B cells and generation of Vav-1 knock - in mice von Kerry Tedford |
| title_full_unstemmed | Characterization of Vav-1 and Vav-2 study of Vav-1 -/- Vav-2 -/- B cells and generation of Vav-1 knock - in mice von Kerry Tedford |
| title_short | Characterization of Vav-1 and Vav-2 |
| title_sort | characterization of vav 1 and vav 2 study of vav 1 vav 2 b cells and generation of vav 1 knock in mice |
| title_sub | study of Vav-1 -/- Vav-2 -/- B cells and generation of Vav-1 knock - in mice |
| topic | Guanosintriphosphatasen (DE-588)4324625-4 gnd Mikroform (DE-588)4039216-8 gnd Molekularbiologie (DE-588)4039983-7 gnd |
| topic_facet | Guanosintriphosphatasen Mikroform Molekularbiologie Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010303062&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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