The gene conversion hypothesis of MHC evolution: studies on the detection of rare segmental exchange variants in the laboratory mouse
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| Format: | Book |
| Language: | English |
| Published: |
2001
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Köln, Univ., Diss., 2000 |
| Physical Description: | 94, VII S. Ill., graph. Darst. : 21 cm |
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| 100 | 1 | |a Sousa, Ana Berta da Fonseca Vieira Álvares e |d 1971- |e Verfasser |0 (DE-588)123098289 |4 aut | |
| 245 | 1 | 0 | |a The gene conversion hypothesis of MHC evolution |b studies on the detection of rare segmental exchange variants in the laboratory mouse |c vorgelegt von Ana Berta da Fonseca Vieira Álvares e Sousa |
| 264 | 1 | |c 2001 | |
| 300 | |a 94, VII S. |b Ill., graph. Darst. : 21 cm | ||
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| 500 | |a Köln, Univ., Diss., 2000 | ||
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| adam_text |
TABLE
OF
CONTENTS
1.
INTRODUCTION
1
1.1.
GENE
CONVERSION
AS
A
MUTATIONAL
MECHANISM
IN
MHC
GENES
1
SELECTION
FOR
DIVERSITY
OPERATES
IN
THE
MHC
1
WHICH
IS
THE
SELECTIVE
MECHANISM
THAT
OPERATES
TO
MAINTAIN
DIVERSITY?
2
WHICH
IS
THE
MOLECULAR
MECHANISM
THAT
OPERATES
TO
GENERATE
DIVERSITY?
3
MOUSE
CLASS
I
GENE
CONVERSION
MUTANTS
4
IN
VITRO
ANALYSIS
OF
MHC
GENE
CONVERSION
MUTATIONS
IN
MAMMALS
5
1.2.
GENE
CONVERSION
AS
A
RECOMBINATIONAL
EVENT
BETWEEN
HOMOLOGOUS
SEQUENCES
8
THE
DOUBLE-STRAND-BREAK
REPAIR
MODEL
8
RECOMBINATION
HAS
BEEN
EXTENSIVELY STUDIED
IN
THE
YEAST
MODEL
SYSTEM
10
HOMOLOGOUS
RECOMBINATION
AND
REPAIR
OF
DSBS
IN
MAMMALIAN
CELLS
14
1.3.
THE
GENE
CONVERSION
HYPOTHESIS
OF
MHC
EVOLUTION
20
2.
MATERIALS
AND
METHODS
23
2.1.
DENATURING
GRADIENT
GEL
ELECTROPHORESIS (DGGE)
23
PRINCIPLE
OF
THE
METHOD
23
CALCULATION
OF
THEORETICAL
MELT
MAPS
BY
COMPUTER
SIMULATION
25
EXPERIMENTAL
DETERMINATION
OF
GRADIENT
BEHAVIOUR
WITH
PERPENDICULAR
GRADIENTS
25
DETECTION
OF
SINGLE
BASE
CHANGES
IN
PARALLEL
GRADIENTS
26
SELECTION
OF
TARGET
REGION
27
DESCRIPTION
OF
TEST
SEQUENCES
28
DGGE
EQUIPMENT
29
ASSORTMENT
OF
TECHNICALITIES
RELEVANT
TO
CASTING
AND
RUNNING
TUBE-DGGS
30
PREPARATION
OF
REAGENTS
FOR
DGGE
31
PREPARATION
OF
(OTHER)
BUFFERS
32
2.2.
ACCESSORY
PROTOCOLS
32
RECOVERY
OF
DNA
FROM
ACRYLAMIDE
TUBE-GELS
BY
CRUSH-SOAK
32
CLEANING
OF
PCR
PRODUCTS WITH EXONUCLEASE
VII
AND
PROTEINASE
K
33
PURIFICATION
OF
PRIMERS BY
ION
EXCHANGE
THROUGH
A
MINIQ
COLUMN
33
CLONING
OF
PCR
PRODUCTS
INTO
PPCR-SCRIPT
AMP
SK(+)
33
PHOTO-INDUCED
CROSS-LINKING
OF
PSORALEN-MODIFIED
PRIMERS
34
LABELLING
OF
RESTRICTED
PBS-TEST
FRAGMENTS
34
IDENTIFICATION
OF
DEPURINATION
AND
DEAMINATION
PRODUCTS
35
3.
RESULTS
36
3.1.
THE
MUTATION
RESOLVING
POTENTIAL
OF
TUBE-DGGE
36
A
K6-GENE
CONVERSION
DETECTOR
36
BEHAVIOUR
OF
TEST
SEQUENCES
IN
PERPENDICULAR
GRADIENT
GELS
37
RESOLUTION
OF
TEST
SEQUENCES
IN
PARALLEL
DGGE
38
DEVELOPMENT
OF
A
TUBE-DGGE
APPARATUS
39
RECOVERY
OF
DNA
FROM
ACRYLAMIDE
TUBE-GELS
BY
CRUSH-SOAK
40
3.2.
PCR-RELATED
BACKGROUND
IN
TUBE-DGGE
41
SELECTION
OF
PFU
POLYMERASE
FOR
AMPLIFICATION
OF
THE
TARGET
REGION
41
OPTIMISATION
OF
PCR
AMPLIFICATION
WITH
PFU
POLYMERASE
42
CLEANING
OF
PCR
PRODUCTS
WITH
EXONUCLEASE
VII
AND
PROTEINASE
K
43
PURIFICATION
AND
ANALYSIS
OF
PRIMER
POPULATIONS
44
PHOTO-INDUCED
CROSS-LINKING
OF
PSORALEN-MODIFIED
PRIMERS
47
3.3.
THE
MUTATION-ENRICHING
POTENTIAL
OF
TUBE-DGGE
48
RECONSTRUCTION
EXPERIMENTS
TO
ASSESS
TUBE-DGGE ENRICHMENT
FACTORS
48
SEMI-QUANTITATIVE
PCR
TO
ASSESS
THE
EXTENT
OF
TRAILING
IN
TUBE-DGGS
49
ESTIMATION
OF
ENRICHMENT
FACTORS
FROM
THE
RUNNING
PROFILE
OF
LABELLED
DNA
49
ESTIMATION
OF
ENRICHMENT
FACTORS
OVER
A
SECOND CYCLE
OF
TUBE-DGGE
51
3.4.
HEAT-INDUCED
BACKGROUND
IN
TUBE-DGGE
53
CONTRIBUTION
OF
DEPURINATION
TO
BACKGROUND
IN
TUBE-DGGS
53
CONTRIBUTION
OF
DEAMINATION
TO
BACKGROUND
IN
TUBE
DGGS
54
SPONTANEOUS
STRAND
CLEAVAGE
AT
ABASIC SITES
55
OPTIMISATION
OF
MELTING
PARAMETERS
IN
THE
DENATURATION-RENATURATION
CYCLE
57
3.5
MUTATION
ENRICHMENT
IN
THE
PCR
PLUS
TUBE-DGGE
COMBINED
PROTOCOL
59
RUNNING
PROFILE
OF
AMPLIFIED
DNA
IN
THE
FIRST
CYCLE
OF
TUBE-DGGE
59
RUNNING
PROFILE
OF
AMPLIFIED
DNA
OVER
A
SECOND
CYCLE
OF
TUBE-DGGE
60
4.
DISCUSSION
63
4.1.
MUTATION
ENRICHMENT
VIA
HETERODUPLEX
RETARDATION
IN
DGGE
63
TUBE-DGGE
IS
NOT
A
POWERFUL
MUTATION-ENRICHING
METHOD
63
SCANNING
METHODS
FOR
DETECTION
OF
UNKNOWN
MUTATIONS
66
GENOTYPIC
SELECTION
OR
DETECTION
OF
RARE
MUTATIONS
68
HIGH-THROUGHPUT
VERSUS
SENSITIVITY
IN
THE DETECTION
OF
RARE
MUTATIONS
71
4.2.
THE GENE
CONVERSION
HYPOTHESIS
OF
MHC
EVOLUTION
REVISITED
72
ADAPTIVE
MUTATORS
IN
THE
IMMUNE
SYSTEM
73
IS
THERE
A
NATURAL
GENE
CONVERSION
MUTATOR
OPERATING
ON
MHC GENES
IN
THE
MOUSE
GERMLINE?
78
THE
STRONG
AND
WEAK
FORMS
OF
THE GENE
CONVERSION
HYPOTHESIS
OF
MHC
EVOLUTION
83
5.
REFERENCES
86
ABSTRACT
I
ZUSAMMENFASSUNG
II
ERKLARUNG
V
ACKNOWLEDGEMENTS
VI
CURRICULUM
VITAE
VII |
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| author | Sousa, Ana Berta da Fonseca Vieira Álvares e 1971- |
| author_GND | (DE-588)123098289 |
| author_facet | Sousa, Ana Berta da Fonseca Vieira Álvares e 1971- |
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| id | DE-604.BV013904755 |
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| indexdate | 2024-08-14T01:00:26Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-009515132 |
| oclc_num | 51997112 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR DE-83 |
| owner_facet | DE-355 DE-BY-UBR DE-83 |
| physical | 94, VII S. Ill., graph. Darst. : 21 cm |
| publishDate | 2001 |
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| spelling | Sousa, Ana Berta da Fonseca Vieira Álvares e 1971- Verfasser (DE-588)123098289 aut The gene conversion hypothesis of MHC evolution studies on the detection of rare segmental exchange variants in the laboratory mouse vorgelegt von Ana Berta da Fonseca Vieira Álvares e Sousa 2001 94, VII S. Ill., graph. Darst. : 21 cm txt rdacontent n rdamedia nc rdacarrier Köln, Univ., Diss., 2000 Maus (DE-588)4169148-9 gnd rswk-swf Genmutation (DE-588)4156638-5 gnd rswk-swf MHC (DE-588)4169760-1 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Maus (DE-588)4169148-9 s MHC (DE-588)4169760-1 s Genmutation (DE-588)4156638-5 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009515132&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Sousa, Ana Berta da Fonseca Vieira Álvares e 1971- The gene conversion hypothesis of MHC evolution studies on the detection of rare segmental exchange variants in the laboratory mouse Maus (DE-588)4169148-9 gnd Genmutation (DE-588)4156638-5 gnd MHC (DE-588)4169760-1 gnd |
| subject_GND | (DE-588)4169148-9 (DE-588)4156638-5 (DE-588)4169760-1 (DE-588)4113937-9 |
| title | The gene conversion hypothesis of MHC evolution studies on the detection of rare segmental exchange variants in the laboratory mouse |
| title_auth | The gene conversion hypothesis of MHC evolution studies on the detection of rare segmental exchange variants in the laboratory mouse |
| title_exact_search | The gene conversion hypothesis of MHC evolution studies on the detection of rare segmental exchange variants in the laboratory mouse |
| title_full | The gene conversion hypothesis of MHC evolution studies on the detection of rare segmental exchange variants in the laboratory mouse vorgelegt von Ana Berta da Fonseca Vieira Álvares e Sousa |
| title_fullStr | The gene conversion hypothesis of MHC evolution studies on the detection of rare segmental exchange variants in the laboratory mouse vorgelegt von Ana Berta da Fonseca Vieira Álvares e Sousa |
| title_full_unstemmed | The gene conversion hypothesis of MHC evolution studies on the detection of rare segmental exchange variants in the laboratory mouse vorgelegt von Ana Berta da Fonseca Vieira Álvares e Sousa |
| title_short | The gene conversion hypothesis of MHC evolution |
| title_sort | the gene conversion hypothesis of mhc evolution studies on the detection of rare segmental exchange variants in the laboratory mouse |
| title_sub | studies on the detection of rare segmental exchange variants in the laboratory mouse |
| topic | Maus (DE-588)4169148-9 gnd Genmutation (DE-588)4156638-5 gnd MHC (DE-588)4169760-1 gnd |
| topic_facet | Maus Genmutation MHC Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009515132&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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