Involvement of the glutathione synthetase gene gsh 2 in phytochelatin biosynthesis of the yeast Schizosaccharomyces pombe:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | German |
| Veröffentlicht: |
1998
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Aachen, Techn. Hochsch., Diss., 1998 |
| Beschreibung: | 111 S. Ill., graph. Darst. |
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| 100 | 1 | |a Lahham, Adnan al- |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Involvement of the glutathione synthetase gene gsh 2 in phytochelatin biosynthesis of the yeast Schizosaccharomyces pombe |c vorgelegt von Adnan Al-Lahham |
| 264 | 1 | |c 1998 | |
| 300 | |a 111 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
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| 500 | |a Aachen, Techn. Hochsch., Diss., 1998 | ||
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| 650 | 0 | 7 | |a Schizosaccharomyces pombe |0 (DE-588)4179645-7 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Phytochelatine |0 (DE-588)4305646-5 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Glutathion-Synthase |0 (DE-588)4386199-4 |2 gnd |9 rswk-swf |
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Datensatz im Suchindex
| _version_ | 1807501071599271936 |
|---|---|
| adam_text |
CONTENTS
PAGE
1.
INTRODUCTION
1.1.
BIOLOGICAL
IMPORTANCE
AND
HAZARDS
OF
HEAVY
METALS
1
1.2.
HEAVY
METAL
RESISTANCE
MECHANISMS
3
1.2.1.
EXTRACELLULAR
COUPLING
3
1.2.2.
METAL
EXCLUSION
4
1.2.3.
BINDING
AT
THE
CELL
WALL
SURFACE
4
1.2.4.
DECREASEDABSORPTION
4
1.2.5.
INCREASED
EFFLUX
5
1.2.6.
VACUOLAR
COMPARTMENTALIZATION
OF
METALS
5
1.2.7.
SYNTHESIS
OF
INTRACELLULAR
CHELATORS
5
1.3.
METALLOTHIONEINS
6
1.4.
GLUTATHIONE
AND
PHYTOCHELATINS;
BIOSYNTHESIS
AND
ROLE
IN
HEAVY
METAL
DETOXIFICATION
8
1.5.
PHYTOCHELATIN
BIOSYNTHESIS
AND
FUNCTIONS
10
1.6.
RESEARCH
OBJECTIVES
15
2.
MATERIALS
2.1.
CHEMICALS
16
2.2.
STRAINS
AND
PLASMIDS
17
2.2.1.
SCHIZOSACCHAROMYCES
POMBE
STRAINS 17
2.2.2.
BACTERIAL
STRAINS
17
2.2.3.
PLASMIDS
17
2.3.
PRIMERS
19
2.4.
MEDIA
PREPARATION
20
3.
METHODS
3.1.
CULTIVATION
OF
ESCHERICHIA
COLL
DH5A
AND
SCHIZOSACCHAROMYCES
POMBE
23
3.2.
BIOCHEMICAL
METHODS
23
3.2.1.
STABILITY
OF
CD
S
-PC
'
MUTANTS
23
3.2.2.
RESPIRATORY
COMPETENCE
TESTING
24
3.2.3.
ENZYMATIC
DETERMINATION
OF
GLUTATHIONE
CONTENT
24
3.2.4.
STUDJYTF
THE
GROWTH
BEHAVIOUR
WITH
AND
WITHOUT
CADMIUM
25
3.2.5.
PREPARATION
OF
THE
S.
POMBE
WILD-TYPE
CULTURES
FOR
OPTIMIZATION
OFPC-SYNTHESIS
26
3.2.6.
LYOPHILIZATION
27
3.2.7.
PREPARATION
OF
CELL
CRUDE
EXTRACTS
FOR
RP-HPLC
ANALYSIS
27
3.2.8.
BRADFORD
PROTEIN
ANALYSIS
27
3.2.9.
ATOMIC
ABSORPTION
SPECTROSCOPY
(AAS)
28
3.3.
GENETICAL
METHODS
28
3.3.1.
MUTATION
OF
S
POMBE
AND
SELECTION
OF
MNNG-RESISTANT
MUTANTS
29
3.3.2.
SELECTION
OF
CD-SENSITIVE
MUTANTS
29
3.3.3.
APPLICATION
OF
A
DROPOUT
METHOD
FOR
AUXOTROPHY
DETERMINATION
29
3.4.
CHROMATOGRAPHICAL
METHODS
30
3.4.1.
POST
DERIVATIZATION
ANALYSIS
BY
RP-HPLC
30
3.4.2.
SEPHADEX
G
50
GEL
FILTRATION
CHROMATOGRAPHY
ASSAY 31
3.5.
MOLECULAR
GENETIC
TECHNIQUES
32
3.5.1.
DNA
SEQUENCING
USING
AN
ALFYY
DNA
SEQUENCER
32
4.
RESULTS
4.1.
PHYTOCHELATIN
OPTIMIZATION
34
4.1.1.
MINIMUM
CONCENTRATION
OF
CDCL
2
NEEDED
TO
INDUCE
PCS
IN
THE
S.
POMBE
WILD-TYPE
34
4.1.2.
PHYTOCHELATIN
CONCENTRATION
IN
THE
STATIONARY
GROWTH
PHASE
OF
THE
WILD-TYPE
35
4.1.3.
PHYTOCHELATIN
CONCENTRATION
IN
THE
LOGARITHMIC
PHASE
OF
GROWTH
OF
THE
WILD-TYPE
36
4.2.
RELATIONSHIP
BETWEEN
PC
AND
GSH
CONCENTRATIONS
37
4.3.
MUTAGENESIS
OF
S.
POMBE
D18,
AND
ISOLATION
OF
PC-DEFICIENT
MUTANTS
38
4.4.
STABILITY
AND
GROWTH
RESPONSE
OF
PC-DEFICIENT
CD-SENSITIVE
MUTANTS
TO
OTHER
METALS
41
4.5.
GROWTH
BEHAVIOUR
OF
THE
PC-DEFICIENT
MUTANT
M379,
WITH
AND
WITHOUT
CADMIUM
COMPARED
TO
THE
WILD-TYPE
42
4.6.
RANDOM
SPORE
ANALYSIS
OF
THE
CROSS
M379
CD
S
-PC*
X
WILD-TYPE
43
4.7.
GROWTH
BEHAVIOUR
OF
CD
S
-PC
'
MUTANT
M379/
8
IN
YDP-MEDIUM
AND
IN
MINIMAL
MEDIUM
44
4.8.
THE
PRESENCE
OF
CADMIUM
BINDING
PEPTIDES
IN
THE
MUTANT
M379/
8
AND
THE
WILD-TYPE
46
4.9.
COMPLEMENTATION
OF
THE
MUTATION
BY
TRANSFORMATION
WITH
A
GENOMIC
LIBRARY
52
4.10.
PLASMID
ANALYSIS
AND
SUBCLONING
53
4.11.
SEQUENCING
STRATEGY
OF
THE
3.7
KB
INSERT
54
4.11.1.
SUBCLONING
OF
ORF
1
AND
ORF2
IN
PUR
19
55
4.11.2.
G
50
CHROMATOGRAPHY
AND
RP-HPLC
DETECTION
OF
PC-BIOSYNTHESIS
EXPRESSED
BY
ORF2
55
4.11.3.
COMPARISON
OF
GSH,
FREE
CYSTEINE,
PCS
AND
THIOL
CONTENTS
IN
THE
TRANSFORMANT
WITH
ORF2,
THE
CD
S
MUTANT
M379/
8
AND
THE
WILD-TYPE
59
4.11.3.1.
IN
COMPLETE
MEDIUM
59
4.11.3.2.
IN
MINIMAL
MEDIUM
62
4.11.3.3.
THIOL
CONTENT
64
4.11.4.
SEQUENCE
AND
MOLECULAR
ANALYSIS
OF
ORF2
65
4.11.5.
MOLECULAR
ANATOMY
OF
THE
PUTATIVE
ORF2
AND
ITS
PROMOTOR
73
4.11.6.
TRANSFORMATION
OF
THE
MUTANT
M379/
8
WITH
THE
TRUNCATED
ORF2
75
4.12.
NUCLEOTIDE
AND
AMINO
ACID
SEQUENCE
OF
ORF
1
76
4.13.
SEARCHING
FOR
OTHER
DNA
FRAGMENTS
CONFERRING
PC-SYNTHESIS
AND
CD-RESISTANCE
79
4.14.
SEARCHING
FOR
OTHER
FRAGMENTS
FROM
THE
WILD-TYPE
GENOMIC
LIBRARY
CONFERRING
OTHER
CD-RESISTANCE
MECHANISMS 81
5.
DISCUSSION
5.1.
OPTIMIZATION
OF
PCS
82
5.2.
OTHER
MECHANISMS
INVOLVED
IN
CD-TOLERANCE
OF
S.
POMBE
84
5.3.
DECISION
TO
CHOOSE
THE
MUTANT
M379
FOR
SEARCHING
OF
THE
COMPLEMENTARY
GENE
86
5.4.
THE
COMPLEMENTING
INSERT
86
5.5.
THE
MUTANT
M379/
8
CONTAINS
ENOUGH
OSH
SUBSTRATE
FOR
THE
ACCUMULATION
OFPCS
89
5.6.
TRANSFORMATION
OF
THE
MUTANT
M379/
8
WITH
THE
TRUNCATED
ORF2
LED
TO
THE
FORMATION
OF
PAPILLARY
COLONIES
90
5.7.
COMPLEMENTATION
OF
THE
MUTANT
M379/
8
BY
TRANSFORMATION
WITH
A
CDNA
CLONE
FROM
ARABIDOPSIS
THALIANA
(HEIM,
1997)
91
5.8.
A
LONG
AND
A
SHORT
TRANSCRIPT
IN
THE
COMPLEMENTARY
GENE
WITH
ORF2
(ROHDE,
1997)
92
5.9.
BIOTECHNOLOGICAL
ASPECTS
RELATED
TO
PHYTOCHELATINS
94
5.10.
NUCLEIC
ACID
SEQUENCE
OF
ORF1
SHARES
SIMILARITIES
WITH
THE
HUMAN
NUCLEIC
ACID
SEQUENCE
94
6.
SUMMARY
AND
CONCLUSIONS
95
7.
APPENDIX
97
7.1.
LIST
OF
ABBREVIATIONS
97
REFERENCES
100 |
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| author | Lahham, Adnan al- |
| author_facet | Lahham, Adnan al- |
| author_role | aut |
| author_sort | Lahham, Adnan al- |
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| bvnumber | BV012192638 |
| classification_tum | CHE 832d BIO 325d BIO 180d |
| ctrlnum | (OCoLC)54430430 (DE-599)BVBBV012192638 |
| discipline | Biologie Chemie |
| format | Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV012192638 |
| illustrated | Illustrated |
| indexdate | 2024-08-16T00:15:46Z |
| institution | BVB |
| language | German |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-008262634 |
| oclc_num | 54430430 |
| open_access_boolean | |
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| owner_facet | DE-M49 DE-BY-TUM DE-355 DE-BY-UBR DE-11 |
| physical | 111 S. Ill., graph. Darst. |
| publishDate | 1998 |
| publishDateSearch | 1998 |
| publishDateSort | 1998 |
| record_format | marc |
| spelling | Lahham, Adnan al- Verfasser aut Involvement of the glutathione synthetase gene gsh 2 in phytochelatin biosynthesis of the yeast Schizosaccharomyces pombe vorgelegt von Adnan Al-Lahham 1998 111 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Aachen, Techn. Hochsch., Diss., 1998 Gen (DE-588)4128987-0 gnd rswk-swf Schizosaccharomyces pombe (DE-588)4179645-7 gnd rswk-swf Phytochelatine (DE-588)4305646-5 gnd rswk-swf Glutathion-Synthase (DE-588)4386199-4 gnd rswk-swf Biosynthese (DE-588)4006902-3 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Schizosaccharomyces pombe (DE-588)4179645-7 s Phytochelatine (DE-588)4305646-5 s Biosynthese (DE-588)4006902-3 s Glutathion-Synthase (DE-588)4386199-4 s Gen (DE-588)4128987-0 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008262634&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Lahham, Adnan al- Involvement of the glutathione synthetase gene gsh 2 in phytochelatin biosynthesis of the yeast Schizosaccharomyces pombe Gen (DE-588)4128987-0 gnd Schizosaccharomyces pombe (DE-588)4179645-7 gnd Phytochelatine (DE-588)4305646-5 gnd Glutathion-Synthase (DE-588)4386199-4 gnd Biosynthese (DE-588)4006902-3 gnd |
| subject_GND | (DE-588)4128987-0 (DE-588)4179645-7 (DE-588)4305646-5 (DE-588)4386199-4 (DE-588)4006902-3 (DE-588)4113937-9 |
| title | Involvement of the glutathione synthetase gene gsh 2 in phytochelatin biosynthesis of the yeast Schizosaccharomyces pombe |
| title_auth | Involvement of the glutathione synthetase gene gsh 2 in phytochelatin biosynthesis of the yeast Schizosaccharomyces pombe |
| title_exact_search | Involvement of the glutathione synthetase gene gsh 2 in phytochelatin biosynthesis of the yeast Schizosaccharomyces pombe |
| title_full | Involvement of the glutathione synthetase gene gsh 2 in phytochelatin biosynthesis of the yeast Schizosaccharomyces pombe vorgelegt von Adnan Al-Lahham |
| title_fullStr | Involvement of the glutathione synthetase gene gsh 2 in phytochelatin biosynthesis of the yeast Schizosaccharomyces pombe vorgelegt von Adnan Al-Lahham |
| title_full_unstemmed | Involvement of the glutathione synthetase gene gsh 2 in phytochelatin biosynthesis of the yeast Schizosaccharomyces pombe vorgelegt von Adnan Al-Lahham |
| title_short | Involvement of the glutathione synthetase gene gsh 2 in phytochelatin biosynthesis of the yeast Schizosaccharomyces pombe |
| title_sort | involvement of the glutathione synthetase gene gsh 2 in phytochelatin biosynthesis of the yeast schizosaccharomyces pombe |
| topic | Gen (DE-588)4128987-0 gnd Schizosaccharomyces pombe (DE-588)4179645-7 gnd Phytochelatine (DE-588)4305646-5 gnd Glutathion-Synthase (DE-588)4386199-4 gnd Biosynthese (DE-588)4006902-3 gnd |
| topic_facet | Gen Schizosaccharomyces pombe Phytochelatine Glutathion-Synthase Biosynthese Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008262634&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT lahhamadnanal involvementoftheglutathionesynthetasegenegsh2inphytochelatinbiosynthesisoftheyeastschizosaccharomycespombe |