Metabolic engineering of the phenylpropanoid pathway: expression of the gene for phenylacrylic acid decarboxylase in plants and study of the effect on plant development
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
1992
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Korrigierte, überarbeitete Fassung. - Oldenburg, Univ., Diss., 1993 |
| Beschreibung: | II, 117 S. Ill., graph. Darst. |
Internformat
MARC
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| 044 | |a gw |c DE | ||
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| 100 | 1 | |a Clausen, Monika |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Metabolic engineering of the phenylpropanoid pathway |b expression of the gene for phenylacrylic acid decarboxylase in plants and study of the effect on plant development |c eingereicht von Monika Clausen |
| 264 | 1 | |c 1992 | |
| 300 | |a II, 117 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Korrigierte, überarbeitete Fassung. - Oldenburg, Univ., Diss., 1993 | ||
| 650 | 0 | 7 | |a Transgene Pflanzen |0 (DE-588)4311513-5 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Phenylpropanstoffwechsel |0 (DE-588)4174252-7 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Phenylpropanstoffwechsel |0 (DE-588)4174252-7 |D s |
| 689 | 0 | 1 | |a Transgene Pflanzen |0 (DE-588)4311513-5 |D s |
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| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-006342902 | |
Datensatz im Suchindex
| _version_ | 1807506230880501760 |
|---|---|
| adam_text |
TABLE
OF
CONTENT
1.
INTRODUCTION
3
2.
MATERIAL
10
2.1.
STRAINS.
PLASMIDS
AND
MEDIA
10
2.1.1.
IN
CLONING
AND
CHARACTERIZATION
OF
THE
PAD
GENE
11
2.1.2.
IN
GENERATION
OF
ANTI-PAD
SERA
11
2.1.3.
IN
PLANT TRANSFORMATION
11
2.2.
DNA
PRIMERS
USED
IN
THIS
WORK
12
2.3.
CHEMICALS
AND
SUPPLIERS
13
3.
CLONING
AND
MOLECULAR
CHARACTERIZATION
OF
THE
PAD
GENE
FROM
S.
CEREVISIAE
14
3.1
INTRODUCTION
14
3.2
METHODS
15
3.2.1.
MOLECULAR
TECHNIQUES
15
3.2.2.
GENETIC
TECHNIQUES
16
3.3.
RESULTS
16
3.3.1.
DETERMINATION
OF
THE
SMALLEST
DNA
FRAGMENT
COMPLEMENTING
THE
MUTANT
16
3.3.2.
MOLECULAR
CHARACTERIZATION
OF
THE
PAD
GENE
19
3.3.2.1.
DETERMINATION
OF
THE
TRANSCRIPT
SIZE
AND
TRANSCRIPTION
DIRECTION
BY
NORTHERN
BLOT
19
3.3.2.2.
SEQUENCING
OF
THE
PAD
GENE
20
3.3.2.3.
DETERMINATION
OF
THE
TRANSCRIPTION
START
SITE
24
3.3.3.
ANALYSIS
OF
THE
PAD
GENE
FUNCTION
25
3.3.3.1
.
MANIPULATION
OF THE
5'
AND
3'
ENDS
OF
THE
PAD
GENE
FOR
FURTHER
CLONING
26
3.3.3.2.
FUNCTIONAL
TEST
TO CONFIRM
THE
PAD
ACTIVITY
OF
THE
CLONED
GENE
26
3.3.3.3.
GENE
REPLACEMENT
28
3.3.3.3.1.
SOUTHERN
BLOT
ANALYSIS
29
3.3.3.3.2.
SEGREGATION
ANALYSIS
31
3.4.
DISCUSSION
33
4.
GENERATION
AND
CHARACTERIZATION
OF
ANTI-PAD
SERUM
39
4.1.
INTRODUCTION
39
4.2.
METHODS
39
4.3.
RESULTS
41
4.3.1.
GENERATION
OF THE
OVEREXPRESSION
CONSTRUCT
41
4.3.2.
DETERMINATION
OF
PAD
MRNA
ACCUMULATION
AFTER
HORMONE
INDUCTION
44
4.3.3.
ACCUMULATION
OF
RECOMBINANT
PROTEIN
AFTER
HORMONE
INDUCTION
44
4.3.4.
DETERMINATION
OF
PAD
ACTIVITY
IN
INDUCED
AND
UNINDUCED
CULTURE
46
4.3.5.
IMMUNIZATION,
CHARACTERIZATION
AND
USE
OF
THE
ANTISERA
TO
DETERMINE
PAD
PROTEIN
IN
PLANTS
47
4.3.5.1.
CHARACTERIZATION
OF
THE
DETECTION
LIMIT
FOR
THE
ANTIGEN
BY
VARIOUS
ANTISERA
47
4.3.5.2.
USE
OF
THE
ANTISERA
TO
DETERMINE
PAD
PROTEINS
IN
PLANTS
48
4.3.6.
CHARACTERIZATION
OF
THE
SERA
48
4.3.6.1.
IN-VITRO
TRANSLATION
AND
IMMUNOPRECIPITATION
49
4.3.7.
WESTERN
BLOT
ANALYSIS
IN
DIFFERENT
YEAST
STRAINS
INCLUDING
THE
PAD::LEU2
REPLACEMENT
51
4.4.
DISCUSSION
52
5.
EXPRESSION
OF
THE
PAD
GENE
IN
PLANTA
AND
INVESTIGATIONS
ON
IT'S
EFFECT
ON
PLANT
DEVELOPMENT
56
5.1.
INTRODUCTION
56
5.2.
METHODS
58
5.3.
RESULTS
60
5.3.1.
ANALYSIS
OF
PAD
HOMOLOGS
IN
PLANTA
60
5.3.2.
COMPARISON
OF THE
CODON
USAGE
FOR
THE
PAD
GENE
WITH
DICOT
PLANTS
61
5.3.3
GENERATION
OF
DIFFERENT
EXPRESSION
CONSTRUCTS
AND
CONFIRMATION
OF
THE
TISSUE
SPECIFICITY
OF
THE
PROMOTERS
62
5.3.4.
BACTERIAL
AND
PLANT
TRANSFORMATION
64
5.3.4.1.
TRANSFORMATIONS
OF
NICOTIANA
TABACUM
(XANTHI)
65
5.3.4.2.
TRANSFORMATION
OF
ARABIDOPSIS
THALIANA
(COLUMBIA)
66
5.3.5.
ANALYSIS
OF
TRANSGENIC
PLANTS
67
5.3.5.1.
GENETIC
DETERMINATION
OF THE
COPY
NUMBER
67
5.3.5.2.
LEVELS
OF
PAD
DNA
OR
RNA
IN
THE
DIFFERENT
TRANGENIC
LINES
69
5.3.6.
OBSERVATION
OF
PHENOTYPIC
CHANGES
72
5.3.6.1
.
OBSERVATIONS
OF
THE
PHENOTYPE
IN
PRIMARY
TRANSFORMANTS
72
5.3.6.2.
CHANGES
OF
PHENOTYPE
IN
SUBSEQUENT
GENERATIONS
AND
ANALYSIS
OF
EFFECTS
ON
PLANT
PHYSIOLOGY
-
74
5.3.7.
DETERMINATION
OF
PHYSIOLOGICAL
CHANGES
IN
TRANSGENIC
PLANTS
80
5.3.7.1
.
COMPARISON
OF
THE
LIGNIFIED
AREA
OF
TRANSGENIC
AND
WILD-TYPE
PLANTS
80
5.3.7.2.
WATER
TRANSPORT
IN
WILD-TYPE
AND
TRANSGENIC
PLANTS
81
5.4.
DISCUSSION
82
6.
SUMMARY
94
ABBREVIATIONS
95
ACKNOWLEDGEMENTS
96
REFERENCES
97 |
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| author | Clausen, Monika |
| author_facet | Clausen, Monika |
| author_role | aut |
| author_sort | Clausen, Monika |
| author_variant | m c mc |
| building | Verbundindex |
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| genre_facet | Hochschulschrift |
| id | DE-604.BV009596709 |
| illustrated | Illustrated |
| indexdate | 2024-08-16T01:37:47Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-006342902 |
| oclc_num | 46241654 |
| open_access_boolean | |
| owner | DE-12 DE-355 DE-BY-UBR DE-11 |
| owner_facet | DE-12 DE-355 DE-BY-UBR DE-11 |
| physical | II, 117 S. Ill., graph. Darst. |
| publishDate | 1992 |
| publishDateSearch | 1992 |
| publishDateSort | 1992 |
| record_format | marc |
| spelling | Clausen, Monika Verfasser aut Metabolic engineering of the phenylpropanoid pathway expression of the gene for phenylacrylic acid decarboxylase in plants and study of the effect on plant development eingereicht von Monika Clausen 1992 II, 117 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Korrigierte, überarbeitete Fassung. - Oldenburg, Univ., Diss., 1993 Transgene Pflanzen (DE-588)4311513-5 gnd rswk-swf Phenylpropanstoffwechsel (DE-588)4174252-7 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Phenylpropanstoffwechsel (DE-588)4174252-7 s Transgene Pflanzen (DE-588)4311513-5 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=006342902&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Clausen, Monika Metabolic engineering of the phenylpropanoid pathway expression of the gene for phenylacrylic acid decarboxylase in plants and study of the effect on plant development Transgene Pflanzen (DE-588)4311513-5 gnd Phenylpropanstoffwechsel (DE-588)4174252-7 gnd |
| subject_GND | (DE-588)4311513-5 (DE-588)4174252-7 (DE-588)4113937-9 |
| title | Metabolic engineering of the phenylpropanoid pathway expression of the gene for phenylacrylic acid decarboxylase in plants and study of the effect on plant development |
| title_auth | Metabolic engineering of the phenylpropanoid pathway expression of the gene for phenylacrylic acid decarboxylase in plants and study of the effect on plant development |
| title_exact_search | Metabolic engineering of the phenylpropanoid pathway expression of the gene for phenylacrylic acid decarboxylase in plants and study of the effect on plant development |
| title_full | Metabolic engineering of the phenylpropanoid pathway expression of the gene for phenylacrylic acid decarboxylase in plants and study of the effect on plant development eingereicht von Monika Clausen |
| title_fullStr | Metabolic engineering of the phenylpropanoid pathway expression of the gene for phenylacrylic acid decarboxylase in plants and study of the effect on plant development eingereicht von Monika Clausen |
| title_full_unstemmed | Metabolic engineering of the phenylpropanoid pathway expression of the gene for phenylacrylic acid decarboxylase in plants and study of the effect on plant development eingereicht von Monika Clausen |
| title_short | Metabolic engineering of the phenylpropanoid pathway |
| title_sort | metabolic engineering of the phenylpropanoid pathway expression of the gene for phenylacrylic acid decarboxylase in plants and study of the effect on plant development |
| title_sub | expression of the gene for phenylacrylic acid decarboxylase in plants and study of the effect on plant development |
| topic | Transgene Pflanzen (DE-588)4311513-5 gnd Phenylpropanstoffwechsel (DE-588)4174252-7 gnd |
| topic_facet | Transgene Pflanzen Phenylpropanstoffwechsel Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=006342902&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT clausenmonika metabolicengineeringofthephenylpropanoidpathwayexpressionofthegeneforphenylacrylicaciddecarboxylaseinplantsandstudyoftheeffectonplantdevelopment |