Identification and characterization of genes with specific expression in dendritic cells:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Regensburg
2002
|
| Schlagworte: | |
| Online-Zugang: | Volltext Volltext Inhaltsverzeichnis |
| Beschreibung: | III, 117 S. Ill., graph. Darst. |
Internformat
MARC
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| 007 | t | ||
| 008 | 030709s2002 ad|| m||| 00||| eng d | ||
| 016 | 7 | |a 968371256 |2 DE-101 | |
| 035 | |a (OCoLC)643409685 | ||
| 035 | |a (DE-599)BVBBV017294293 | ||
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| 100 | 1 | |a Heinz, Sven |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Identification and characterization of genes with specific expression in dendritic cells |c vorgelegt von Sven Heinz |
| 264 | 1 | |a Regensburg |c 2002 | |
| 300 | |a III, 117 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Universität Regensburg |d 2002 | ||
| 650 | 0 | 7 | |a Dendritische Zelle |0 (DE-588)4313846-9 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Genexpression |0 (DE-588)4020136-3 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Dendritische Zelle |0 (DE-588)4313846-9 |D s |
| 689 | 0 | 1 | |a Genexpression |0 (DE-588)4020136-3 |D s |
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| 776 | 1 | 8 | |i Erscheint auch als |n Online-Ausgabe |o urn:nbn:de:bvb:355-opus-2086 |
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| 856 | 4 | 1 | |u https://epub.uni-regensburg.de/10104/ |x Verlag |z kostenfrei |3 Volltext |
| 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=010425959&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 912 | |a ebook | ||
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-010425959 | |
Datensatz im Suchindex
| _version_ | 1807504002061959168 |
|---|---|
| adam_text |
TABLE
OF
CONTENTS
1
INTRODUCTION
.
1
1.1
THE
IMMUNE
SYSTEM
.
1
1.2
DENDRITIC
CELLS:
ANTIGEN
PRESENTING
CELLS
BRIDGING
INNATE
AND
ADAPTIVE
IMMUNITY
.
2
1.2.1
DENDRITIC
CELL
ONTOGENY
.
2
1.2.2
DC
MODEL
SYSTEMS
.
5
1.3
DENDRITIC
CELL
FUNCTION
.
5
1.3.1
ANTIGEN
UPTAKE
.
6
1.3.2
ANTIGEN
PROCESSING
AND
PRESENTATION
.
7
1.3.3
COSTIMULATION
.
8
1.3.4
HELPER
T
CELL
POLARIZATION
.
9
1.3.5
TOLERANCE
INDUCTION
.
10
1.3.6
INTERACTIONS
WITH
OTHER
CELLS
OF
THE
IMMUNE
SYSTEM
.
10
1.4
DC
MORPHOLOGY
AND
FUNCTION
AT
THE
DIFFERENT
STAGES
OF
DIFFERENTIATION
.
11
1.4.1
CHARACTERISTIC
MOLECULES
.
13
2
RESEARCH
OBJECTIVES
.
15
3
MATERIALS
AND
METHODS
.
17
3.1
EQUIPMENT
AND
MATERIALS
.
17
3.1.1
EQUIPMENT
.
17
3.1.2
MATERIALS
.
17
3.1.3
CHEMICALS
.
18
3.1.4
DNA
OLIGONUCLEOTIDES
.
18
3.1.5
ANTIBODIES
.
19
3.1.6
ENZYMES,
INHIBITORS
AND
KITS
.
19
3.1.7
MOLECULAR
WEIGHT
STANDARDS
.
20
3.1.8
PRIMARY
CELLS
AND
CELL
LINES
.
20
3.1.9
BACTERIAL
E.COLI
STRAINS
.
20
3.1.10
PLASMID
VECTORS
.
21
3.2
CELL
ISOLATION
.
21
3.2.1
MONOCYTES
.
21
3.2.2
BLOOD
DENDRITIC
CELL
ISOLATION
BY
MACS
.
22
3.2.3
ISOLATION
OF
GRANULOCYTES
FROM
BUFFY
COATS
.
22
3.3
CELL
CULTURE
.
23
3.3.1
CELL
CULTURE
CONDITIONS
AND
PASSAGING
.
23
3.3.2
ASSESSING
CELL
VITALITY
BY
TRYPAN
BLUE
EXCLUSION
.
23
3.3.3
FREEZING
AND
THAWING
CELLS
.
24
3.3.4
PRIMARY
CELLS
.
24
3.3.4.1
DENDRITIC
CELLS
.
24
3.3.4.2
MACROPHAGES
.
24
3.3.5
CELL
LINES
.
25
3.3.6
MYCOPLASMA
ASSAY
.
25
3.3.7
MIXED
LEUKOCYTE
CULTURE
.
25
3.4
DNA
.
27
3.4.1
TRANSIENT
TRANSFECTION
OF
THP-1
CELLS
WITH
DEAE-DEXTRAN
.
27
TABLE
OF
CONTENTS
3.4.2
AGAROSE
GEL
ELECTROPHORESIS
.
28
3.4.3
DENATURING
ALKALINE
AGAROSE
GELS
FOR
ANALYSIS
OF
SINGLE-STRANDED
DNA
.
29
3.4.4
PURIFICATION
OF
DNA
FRAGMENTS
BY
GEL
EXTRACTION
.
30
3.4.5
REPRESENTATION
DIFFERENCE
ANALYSIS
.
30
3.4.5.1
CDNA-SYNTHESIS
.
31
3.4.5.2
GENERATION
OF
REPRESENTATIONS
.
31
3.4.5.3
DIFFERENCE
ANALYSIS
.
33
3.4.6
REVERSE
DOT
BLOT
.
35
3.4.7
PCR
.
36
3.4.8
RT-PCR
.
37
3.4.8.1
SMART-RACE
.
38
3.4.9
PRECIPITATION
OF
DNA
USING
PEG
.
38
3.4.10
PCR-BASED
SITE-SPECIFIC
MUTAGENESIS
.
39
3.4.11
GENOME
WALKING
.
40
3.4.12
PREPARATION
OF
GENOMIC
DNA
AND
BISULFITE
SEQUENCING
.
41
3.4.13
DNA
SEQUENCING
AND
SEQUENCE
ANALYSIS
.
42
3.5
RNA
.
42
3.5.1
RNA
ISOLATION
BY
ADD
GUANIDINIUM
THIOCYANATE-PHENOL-CHLOROFORM
EXTRACTION
.
42
3.5.2
CSCI
PURIFICATION
OF
RNA
.
44
3.5.3
POLY-A
MRNA
ISOLATION
.
45
3.5.4
ELECTROPHORESIS
OF
RNA
IN
DENATURING
FORMALDEHYDE
AGAROSE
GELS
.
45
3.5.5
NORTHERN
BLOT
-
RNA
TRANSFER
.
45
3.5.6
RADIOACTIVE
LABELING
OF
DNA
.
46
3.5.7
NORTHERN
BLOT
HYBRIDIZATION
.
47
3.6
MOLECULAR
CLONING
.
48
3.6.1
BACTERIAL
CULTURE
.
48
3.6.2
PREPARATION
OF
CHEMICALLY
COMPETENT
E.COLI
.
49
3.6.3
TRANSFORMATION
OF
CHEMICALLY
COMPETENT
E.COLI
.
49
3.6.4
CLONING
.
50
3.6.5
PLASMID
DNA
PREPARATION
.
51
3.7
PROTEIN
METHODS
.
51
3.7.1
NUCLEAR
EXTRACTION
PROCEDURE
.
51
3.7.2
BCA
PROTEIN
ASSAY
.
52
3.7.3
ELECTROPHORETIC
MOBILITY
SHIFT
ASSAY
.
52
3.7.4
IMMUNOPRECIPITATION
.
55
3.7.5
DISCONTINUOUS
SDS-PAGE
.
55
3.7.6
WESTERN
BLOT
.
57
3.7.7
IMMUNOSTAINING
OF
BLOTTED
PROTEINS
.
57
3.7.8
FLOW
CYTOMETRY
.
58
4
RESULTS
.
59
4.1
IDENTIFICATION
OF
GENES
WITH
DC-SPECIFIC
EXPRESSION
BY
REPRESENTATIONAL
DIFFERENCE
ANALYSIS
.
59
4.2
EXPRESSION
ANALYSES
BY
NORTHERN
BLOT
HYBRIDIZATION
.
62
4.3
MCP-4
.
64
4.3.1
ANALYSIS
OF
MCP-4
EXPRESSION
DURING
DC
DIFFERENTIATION
.
64
4.3.2
BLOOD
DC
PRODUCE
MCP-4
MRNA
.
66
TABLE
OF
CONTENTS
4.3.3
DCS
UTILIZE
THE
SAME
MCP-4
PROMOTER
AS
DERMAL
FIBROBLASTS
.
67
4.3.4
DIFFERENT
REGULATORY
PATHWAYS
GOVERN
MCP-4
EXPRESSION
IN
DCS
AND
DERMAL
FIBROBLASTS
.
67
4.3.5
CPG
METHYLATION
ANALYSIS
OF
THE
PROXIMAL
MCP-4
PROMOTER
.
68
4.3.6
DENDRITIC
CELL-SPECIFIC
DEMETHYLATION
OF
THE
MCP-4
PROMOTER
ALLOWS
BINDING
OF
A
NUCLEAR
FACTOR
.
71
4.3.7
NUCLEAR
FACTOR
BINDING
AT
THE
-80
BP
CPG
IS
NECESSARY
FOR
MAXIMAL
MCP-4
PROMOTER
ACTIVITY
IN
REPORTER
ASSAYS
IN
TRANSIENTLY
TRANSFECTED
THP-1
CELLS
.
74
4.3.8
CPG
DEMETHYLATION
OF
THE
MCP-4
PROMOTER
DURING
MONOCYTE
TO
DENDRITIC
CELL
DIFFERENTIATION
IS
DIFFERENTIATION
STAGE-DEPENDENT
.
75
4.4
HEP27
.
77
4.4.1
PREDOMINANT
EXPRESSION
OF
HEP27
IN
MONOCYTE-DERIVED
DENDRITIC
CELLS
.
77
4.4.2
TISSUE
EXPRESSION
OF
HEP27
MRNA
.
78
4.4.3
DC
AND
HEPG2
CELLS
UTILIZE
DIFFERENT
HEP27
PROMOTERS
.
80
4.4.4
GENOMIC
ORGANIZATION
OF
HEP27
.
81
4.4.5
EXCLUSIVE
UTILIZATION
OF
THE
UPSTREAM
HEP27
PROMOTER
BY
MONOCYTE-DERIVED
DENDRITIC
CELLS
AND
ALTERNATIVE
SPLICING
.
82
4.4.6
BUTYRATE
TREATMENT
ACTIVATES THE
DOWNSTREAM
HEP27
PROMOTER
.
84
5
DISCUSSION
.
87
5.1
IDENTIFICATION
OF
GENES
WITH
DC-SPECIFIC
EXPRESSION
.
87
5.2
MCP-4
.
87
5.3
HEP27
.
97
5.4
COMPLEMENT
C1Q
C-CHAIN
.
99
5.5
15-LIPOXYGENASE
.
101
5.6
FOLATE
RECEPTOR
P
.
103
6
SUMMARY
.
105
7
REFERENCES
.
107 |
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| author_facet | Heinz, Sven |
| author_role | aut |
| author_sort | Heinz, Sven |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV017294293 |
| illustrated | Illustrated |
| indexdate | 2024-08-16T01:02:23Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-010425959 |
| oclc_num | 643409685 |
| open_access_boolean | 1 |
| owner | DE-355 DE-BY-UBR DE-12 DE-384 DE-473 DE-BY-UBG DE-703 DE-1051 DE-824 DE-29 DE-91 DE-BY-TUM DE-19 DE-BY-UBM DE-1049 DE-92 DE-739 DE-20 |
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| physical | III, 117 S. Ill., graph. Darst. |
| psigel | ebook |
| publishDate | 2002 |
| publishDateSearch | 2002 |
| publishDateSort | 2002 |
| record_format | marc |
| spelling | Heinz, Sven Verfasser aut Identification and characterization of genes with specific expression in dendritic cells vorgelegt von Sven Heinz Regensburg 2002 III, 117 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Dissertation Universität Regensburg 2002 Dendritische Zelle (DE-588)4313846-9 gnd rswk-swf Genexpression (DE-588)4020136-3 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Dendritische Zelle (DE-588)4313846-9 s Genexpression (DE-588)4020136-3 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:355-opus-2086 https://nbn-resolving.org/urn:nbn:de:bvb:355-opus-2086 Resolving-System kostenfrei Volltext https://epub.uni-regensburg.de/10104/ Verlag kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010425959&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Heinz, Sven Identification and characterization of genes with specific expression in dendritic cells Dendritische Zelle (DE-588)4313846-9 gnd Genexpression (DE-588)4020136-3 gnd |
| subject_GND | (DE-588)4313846-9 (DE-588)4020136-3 (DE-588)4113937-9 |
| title | Identification and characterization of genes with specific expression in dendritic cells |
| title_auth | Identification and characterization of genes with specific expression in dendritic cells |
| title_exact_search | Identification and characterization of genes with specific expression in dendritic cells |
| title_full | Identification and characterization of genes with specific expression in dendritic cells vorgelegt von Sven Heinz |
| title_fullStr | Identification and characterization of genes with specific expression in dendritic cells vorgelegt von Sven Heinz |
| title_full_unstemmed | Identification and characterization of genes with specific expression in dendritic cells vorgelegt von Sven Heinz |
| title_short | Identification and characterization of genes with specific expression in dendritic cells |
| title_sort | identification and characterization of genes with specific expression in dendritic cells |
| topic | Dendritische Zelle (DE-588)4313846-9 gnd Genexpression (DE-588)4020136-3 gnd |
| topic_facet | Dendritische Zelle Genexpression Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:355-opus-2086 https://epub.uni-regensburg.de/10104/ http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010425959&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT heinzsven identificationandcharacterizationofgeneswithspecificexpressionindendriticcells |