The regulation of endosperm-specific gene expression in maize:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | German |
| Veröffentlicht: |
1999
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | V, 120 S. Ill., graph. Darst. |
Internformat
MARC
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| 001 | BV012887616 | ||
| 003 | DE-604 | ||
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| 049 | |a DE-355 |a DE-83 |a DE-188 | ||
| 100 | 1 | |a Guo, Yan |d 1967- |e Verfasser |0 (DE-588)121483622 |4 aut | |
| 245 | 1 | 0 | |a The regulation of endosperm-specific gene expression in maize |c Yan Guo |
| 264 | 1 | |c 1999 | |
| 300 | |a V, 120 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a Köln, Univ., Diss., 1999 | ||
| 650 | 0 | 7 | |a Mais |0 (DE-588)4037135-9 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Genregulation |0 (DE-588)4122166-7 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Endosperm |0 (DE-588)4367419-7 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Mais |0 (DE-588)4037135-9 |D s |
| 689 | 0 | 1 | |a Endosperm |0 (DE-588)4367419-7 |D s |
| 689 | 0 | 2 | |a Genregulation |0 (DE-588)4122166-7 |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=008772231&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-008772231 | |
Datensatz im Suchindex
| _version_ | 1807683450646298624 |
|---|---|
| adam_text |
INDEX
I
CONTENTS
1.
INTRODUCTION
1
1.1
.OCCURANCE
AND
DIFFERENTIATION
OF
THE
ENDOSPERM
.
1
1.2.
EFFECTS
OF
PHYTOHORMONES
IN
MAIZE
ENDOSPERM
DEVELOPMENT
.
3
1.3
REGULATION
OF
GENE
EXPRESSION
IN
THE
ENDOSPERM
.
3
1.3.1.
CENTRAL
ENDOSPERM
DOMAIN
.
3
1.3.2.
ALEURONE
DOMAIN
.
4
1.3.3.
BASAL
LAYER
DOMAIN
.
6
1.4
SUGAR
TRANSPORT
AND
ACCUMULATION
IN
MAIZE
ENDOSPERM
.
8
1.5
PLANT
TRANSFER
CELLS
.
11
1.5.1
LOCATION
OF
TRANSFER
CELLS
.
11
1.5.2
MAIZE
ENDOSPERM
TRANSFER
CELL
.
12
2.MATERIALS
AND
METHODS
.
14
2.1.
ABBREVIATIONS
.
14
2.2.
MATERIALS
.
15
2.2.1.
PLANT
MATERIALS
.
15
2.2.2.
BACTERIAL
STRAINS,
YEAST
STRAINS
AND
CLONING
VECTORS
.
15
2.2.3.
CHEMICAL
AND
ENZYME
.
16
2.2.4.
MEDIA
.
16
2.2.5.
BUFFERS
AND
SOLUTIONS
.
16
2.3.
METHODS
.
19
2.3.1
PLASMID
DNA
EXTRACTION
.
19
2.3.2
ISOLATION
OF
PLANT
GENOMIC
DNA
.
19
2.3.3
ISOLATION
OF
PLANT
TOTAL
RNA
AND
POLY
(A)
+
RNA
.
19
2.3.4
GENOMIC
SOUTHERN
BLOTTING
ANALYSIS
.
20
2.3.5
RNA
NORTHERN
BLOTTING
ANALYSIS
.
20
2.3.6
PREPARATION
OF
|A-32PJ-LABELLED
PROBES
.
21
2.3.6.1
RANDOM
PRIME
LABELLED
PROBES
.
21
2.3.6.2
PROBE
LABELLED
BY
END-FILLING
.
21
INDEX
U
2.3.6.3
CDNA
PROBE
LABELLED
.
21
2.3.7
PROTEIN
WESTERN
BLOTTING
ANALYSIS
.
22
2.3.8
PCR
AMPLIFICATION
ANALYSIS
.
23
2.3.8.1
GENOMIC
DNA
ORCDNA
FIRST
STRAND
PCR
AMPLIFICATION
.
23
2.3.8.2
PLASMID
DNA
PCR
AMPLIFICATION
.
23
2.3.8.3
PCR
AMPLIFICATION
ASSAYS
TRANSFORMED
CALLI
OR
PLANTS
.
24
2.3.9
GUS
ACTIVITY
ASSAY
.
24
2.3.9.1
HISTOCHEMICAL
DETECTION
OF
GUS
ACTIVITY
.
24
2.3.9.2
QUANTITATIVE
ASSAY
OF
GUS
ACTIVITY
.
24
2.3.10
EMSA
(ELECTROPHORETIC
MOBILITY
SHIFT
ASSAY)
.
25
2.3.10.1
CRUDE
NUCLEAR
EXTRACT
ISOLATION
FROM
MAIZE
ENDOSPERM
.
25
2.3.10.2
EMSA
REACTION
.
25
2.3.11
TRANSIENT
GENE
EXPRESSION
IN
TOBACCO
PROTOPLASTS
VIA
PEG
TRANSFORMATION
25
2.3.11.1
PROTOPLAST
PREPARATION
.
25
2.3.11.2
PROTOPLAST
TRANSFORMATION
.
26
2.3.11.3
GUS
ACTIVITY
ASSAY
.
26
2.3.12
PLANT
TRANSFORMATION
.
26
2.3.12.1
MAIZE
TRANSFORMATION
.
26
2.3.12.2
TOBACCO
TRANSFORMATION
.
26
2.3.12.3
ARABIDOPSIS
TRANSFORMATION
AGROBACTERIUM
VACUUM
INFILTRATION.
27
2.3.13
DIFFERENTIAL
DISPLAY
RT-PCR
.
28
2.3.13.1
REVERSE
TRANSCRIPTION
.
28
2.3.13.2
PCR
AMPLIFICATION
.
28
2.3.13.3
CLONING
DDRT-PCR
PRODUCTS
.
29
3.
RESULTS
.
30
3.1
BETL-2
PROMOTER
ANALYSIS
IN
TRANSFORMED
MAIZE
PLANTS
.
30
3.1.1
CONSTRUCTION
OF
THE
BETL-2
PROMOTER/GUS
(BETL-2/GUS)
FUSION
PLASMID
30
3.1.2.
TRANSFORMATION
OF
THE
BETL-2
PROMOTER/GUS
(BETL-2/GVS)
PLASMID
INTO
MAIZE
EMBRYOGENIC
PROTOPLASTS
.
31
3.1.3.
REGENERATION
AND
ANALYSIS
OF
TO
PLANTS
.
32
3.1.4
ANALYSIS
OF
T,
AND
T2
TRANSGENIC
PLANTS
.
34
INDEX
HI
3.1.5
WESTERN
BLOT
ANALYSIS
OF
BETL-2/GGS
TRANSFORMED
MAIZE
PLANTS
.
35
3.1.6
DEVELOPMENTAL
EXPRESSION
OF
THE
GUS
GENE
IN
BE//-2/GUS
TRANSFORMED
PLANTS
.
38
3.2
BETL-2
PROMOTER
ACTIVITY
IN
TRANSFORMED
TOBACCO
PLANTS
.
41
3.2.1
TISSUE-SPECIFIC
GUS
EXPRESSION
OF
THE
5'
UNILATERAL
BETL-2
PROMOTER/GUS
DELETION
CONSTRUCTS
.
42
3.2.1.1
CONSTRUCTION
OF
THE
5
'
UNILATERAL
BETL-2
PROMOTER/GUS
DELETIONS
.
42
3.2.1.2
TOBACCO
TRANSFORMATION
AND
ANALYSIS
OF
GUS
EXPRESSION
.
43
3.2.2
TISSUE-SPECIFIC
GUS
EXPRESSION
OF
THE
BETL-2
PROMOTER
INTERNAL
DELETION
/
GUS
CONSTRUCTS
.
45
3.2.2.1
CONSTRUCTION
OF
THE
BETL-2
PROMOTER
INTERNAL
DELETION/GUS
PLASMIDS.
45
3.2.2.2
GUS
ACTIVITY
OF
THE
INTERNAL
DELETION
CONSTRUCTS
IN
TRANSFORMED
TOBACCO
PLANTS
.
47
3.2.3
GUS
ACTIVITY
WAS
SPECIFICALLY
FOUND
IN
MATERNAL
TISSUE
OF
TRANSGENIC
TOBACCO
PLANTS
.
78
3.3
ACTIVITY
OF
THE
BETL-2
PROMOTER/GUS
(BETL-2/GUS)
CONSTRUCT
IN
ARABIDOPSIS.50
3.3.1
TISSUE-SPECIFIC
(GUS)
EXPRESSION
OF
BETL-2
/GUS
DELETION
CONSTRUCTS
IN
ARABIDOPSIS
.
50
3.3.2
THE
EFFECT
ON
BETL-2IGGS
ACTIVITY
IN
ARABIDOPSIS
TESTA
MUTANTS
.
51
3.4
ELECTROPHORETIC
MOBILITY
SHIFT
ASSAY
(EMSA)
ANALYSIS
OF
THE
BETL-2
PROMOTER
54
3.4.1.
TWO
MOTIFS,
GATA
AND
GTATTAATTC,
INTERACT
SPECIFICALLY
WITH
NUCLEAR
PROTEIN
FROM
UPPER
AND
BASAL
PART
ENDOSPERM
NUCLEAR
PROTEIN
EXTRACTS
55
3.4.1.1
"
GATA
"
AS
A
CORE
MOTIF
INTERACTS
WITH
BOTH
THE
MAIZE
ENDOSPERM
UPPER
AND
BASAL
PART
NUCLEAR
EXTRACTS
.
55
3.4.1.2
A
"
GTATTAATTC
"
MOTIF
INTERACTS
WITH
MAIZE
BASAL
PART
ENDOSPERM
NUCLEAR
EXTRACTS
.
57
3.5
A
MYB-LIKE
TRANSCRIPTION
FACTOR,
MZMRP-1,
INTERACTS
WITH
THE
BETL-2
PROMOTER
IN
TOBACCO
PROTOPLASTS
.
60
3.5.1
CONSTRUCTION
OF
THE
MZMRP-L
/PRT105
EXPRESSION
VECTOR
.
60
3.5.2
INTERACTION
OF
MZMRP-1
WITH
THE
BETL-2
PROMOTER
IN
TOBACCO
PROTOPLASTS.
61
INDEX
IV
3.6
ANALYSIS
OF
ANTISENSE
BETL-2
CDNA
TRANSFORMED
MAIZE
PLANTS
.
63
3.6.1
CONSTRUCTION
OF
ANTISENSE
BETL-2
EXPRESSION
PLASMID
.
63
3.6.2
TRANSFORMATION
OF
THE
ANTISENSE
BETL-2
CDNA
CONSTRUCT
INTO
MAIZE
.
63
3.6.3
SOUTHERN
BLOT
ANALYSIS
OF
THE
CHIMERIC
ANTISENSE
BETL-2
GENE
IN
MAIZE
TRANSFORMANTS
64
3.6.4
RT-PCR
ANALYSIS
OF
THE
T
YY
ANTISENSE
BETL-2
PLANTS
.
65
3.6.5
NORTHERN
BLOT
ANALYSIS
OF
THE
T
YY
ANTISENSE
BETL-2
TRANSFORMED
PLANTS
.
66
3.6.6
WESTERN
BLOT
ANALYSIS
OF
A
SINGLE
KERNEL
FROM
THE
ANTISENSE
BETL-2
TRANSFORMED
PLANTS
.
67
3.7.
COMPARISION
OF
GENES
EXPRESSED
IN
THE
ENDOSPERM
DEFECTIVE
MUTANT
AND
WILD
TYPE
MAIZE
BY
DDRT-PCR
.
69
3.7.1
CHARACTERISTICS
OF
THE
MAIZE
ENDOSPERM
DEFECTIVE
MUTANT
DE-BL8
.
&)
3.7.2
DIFFERENTIAL
DISPLAY
MRNA
BASED
ON
PCR
AMPLIFICATION
.
69
3.7.3
DDRT-PCR
ANALYSIS
OF
WILD-TYPE
B37
AND
MUTANT
DE-B18
.
71
3.7.4
ANALYSIS
OF
GENES
ISOLATED
BY
DIFFERENTIAL
DISPLAY
.
78
3.7.4.1
REVERSE
NORTHERN
BLOT
AND
NORTHERN
BLOT
ANALYSIS
.
78
3.7.4.2
PCR
AMPLIFICATION
AND
RFLP
ANALYSIS
.
80
3.7.4.2.1
PCR
ANALYSIS
OF
DDRT-PCR
CLONES
.
80
3.7.4.2.2
RFLP
ANALYSIS
OF
DDRT-PCR
CLONES
.
82
3.7.4.3
MAPPING
OF
THE
DE-BL8
LOCUS
.
85
3.7.4.4
DISTANCE
OF
DD-28
TO
DE-B18
LOCUS
.
86
4.
DISCUSSION
.
88
4.1.
CHARACTERISTICS
OF
THE
BETL-2
PROMOTER
.
88
4.1.1.
PROMOTER-REPORTER
GENE
EXPRESSION
IN
MAIZE
.
88
4.1.2.
DIFFERENT
LEVELS
OF
OUS
ACTIVITY WERE
OBTAINED
FROM
INDEPENDENT
TRANSFORMED
MAIZE
PLANTS
.
89
4.1.3.
HETEROLOGOUS
TRANSGENIC
EXPRESSION
OF
THE
BETL-2
/GUS
IN
TOBACCO
AND
ARABIDOPSIS
.
90
4.1.3.1
A
165BP
REGION
OF
THE
BETL-2
PROMOTER
IS
SUFFICIENT
TO
DIRECT
BETL-2
GENE
EXPRESSION
.
91
4.1.3.2.
MATEMAL-TISSUE-SPECIFIC
EXPRESSION
OF
GUS
IN
BETL-2/GUS
TRANS
FORMED
TOBACCO
PLANTS
.
92
INDEX
V
4.1.3.4.
GUS
ACTIVITY
OF
THE
BETL-2/G\IS
TRANSFORMED
ARABIDOPSIS
PLANTS
WAS
AFFECTED
BY
THREE
TESTA
MUTATIONS,
TTL-1,
TT4-L
AND
TTG-1
.
92
4.1.4.
TWO
C/5-ACTING
MOTIFS
PRESENT
IN
THE
BETL-2
PROMOTER
ARE
SPECIFICALLY
BOUND
BY
FACTORS
PRESENT
IN
MAIZE
BASAL
ENDOSPERM
NUCLEAR
EXTRACTS
.
93
4.1.5
A
MAIZE
ENDOSPERM
BASAL
LAYER
SPECIFIC
MYB
TRANSCRIPTION
FACTOR
INTERACTS
WITH
THE
BETL-2
PROMOTER
.
94
4.1.6
A
MODEL
TO
SHOW
THE
PUTATIVE
REGULATORY
ELEMENTS
WITHIN
THE
BETL-2
PROMOTER
.
95
4.2.
CHARACTERISATION
OF
THE
ANTISENSE
BETL-2
TRANSGENIC
MAIZE
PLANTS
.
95
4.2.1.
INTEGRATION
AND
EXPRESSION
OF
THE
ANTISENSE
BETL-2
CDNA
INTO
MAIZE
.
95
4.2.2.
REDUCED
EXPRESSION
OF
THE
BETL-2
GENE
DOES
NOT
AFFECT
THE
PHENOTYPE
OF
TRANSFORMED
PLANTS
.
97
4.3.
ISOLATION
OF
DIFFERENTIAL
EXPRESSED
GENES
BETWEEN
MAIZE
ENDOSPERM
DEFECTIVE
MUTANT
(DE-B18)
AND
ITS
WILD-TYPE
B37
.
97
4.3.1.
CLONING
GENES
EXPRESSED
IN
WILD-TYPE
BUT
NOT
IN
DE-B18
MAIZE
BY
DDRT
PCR
.
98
4.3.2.
GENES
DIFFERENTLY
EXPRESSED
BETWEEN
WILD-TYPE
AND
MUTANT
.
99
4.3.3.
GENETIC
MAPPING
OF
THE
DE-B18
LOCUS
.
101
5.
SUMMARY
.
103
6.
REFERENCES.
107 |
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| author_GND | (DE-588)121483622 |
| author_facet | Guo, Yan 1967- |
| author_role | aut |
| author_sort | Guo, Yan 1967- |
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| bvnumber | BV012887616 |
| ctrlnum | (OCoLC)64648232 (DE-599)BVBBV012887616 |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV012887616 |
| illustrated | Illustrated |
| indexdate | 2024-08-18T00:34:37Z |
| institution | BVB |
| language | German |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-008772231 |
| oclc_num | 64648232 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR DE-83 DE-188 |
| owner_facet | DE-355 DE-BY-UBR DE-83 DE-188 |
| physical | V, 120 S. Ill., graph. Darst. |
| publishDate | 1999 |
| publishDateSearch | 1999 |
| publishDateSort | 1999 |
| record_format | marc |
| spelling | Guo, Yan 1967- Verfasser (DE-588)121483622 aut The regulation of endosperm-specific gene expression in maize Yan Guo 1999 V, 120 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Köln, Univ., Diss., 1999 Mais (DE-588)4037135-9 gnd rswk-swf Genregulation (DE-588)4122166-7 gnd rswk-swf Endosperm (DE-588)4367419-7 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Mais (DE-588)4037135-9 s Endosperm (DE-588)4367419-7 s Genregulation (DE-588)4122166-7 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008772231&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Guo, Yan 1967- The regulation of endosperm-specific gene expression in maize Mais (DE-588)4037135-9 gnd Genregulation (DE-588)4122166-7 gnd Endosperm (DE-588)4367419-7 gnd |
| subject_GND | (DE-588)4037135-9 (DE-588)4122166-7 (DE-588)4367419-7 (DE-588)4113937-9 |
| title | The regulation of endosperm-specific gene expression in maize |
| title_auth | The regulation of endosperm-specific gene expression in maize |
| title_exact_search | The regulation of endosperm-specific gene expression in maize |
| title_full | The regulation of endosperm-specific gene expression in maize Yan Guo |
| title_fullStr | The regulation of endosperm-specific gene expression in maize Yan Guo |
| title_full_unstemmed | The regulation of endosperm-specific gene expression in maize Yan Guo |
| title_short | The regulation of endosperm-specific gene expression in maize |
| title_sort | the regulation of endosperm specific gene expression in maize |
| topic | Mais (DE-588)4037135-9 gnd Genregulation (DE-588)4122166-7 gnd Endosperm (DE-588)4367419-7 gnd |
| topic_facet | Mais Genregulation Endosperm Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008772231&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT guoyan theregulationofendospermspecificgeneexpressioninmaize |