Carbon metabolism in Streptomyces coelicolor: a molecular view to glucose kinase and its role in carbon regulation and glucose transport
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| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English German |
| Published: |
2001
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Item Description: | Erlangen-Nürnberg, Univ., Diss., 2001. - Enth. Zsfassung in dt. Sprache |
| Physical Description: | III, 100 S. Ill., graph. Darst. |
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MARC
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| 001 | BV014219249 | ||
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| 100 | 1 | |a Mahr, Kerstin |d 1973- |e Verfasser |0 (DE-588)123554179 |4 aut | |
| 245 | 1 | 0 | |a Carbon metabolism in Streptomyces coelicolor |b a molecular view to glucose kinase and its role in carbon regulation and glucose transport |c von Kerstin Mahr |
| 264 | 1 | |c 2001 | |
| 300 | |a III, 100 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Erlangen-Nürnberg, Univ., Diss., 2001. - Enth. Zsfassung in dt. Sprache | ||
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Record in the Search Index
| _version_ | 1807772509223780352 |
|---|---|
| adam_text |
INDEX
I
1
ZUSAMMENFASSUNG
/
SUMMARY
.
1
2
INTRODUCTION
.
.
.
YY2
2.1
THE
ANTIBIOTIC
PRODUCER
STREPTOMYCES
COETICOTOR.
A
DIFFERENTIATING
PROCARYOTE
_
.
-
2
2
J
CARBOHYDRATE
UPTAKE
IN
SRREPROMYCES.^.^.
.
2.3
CARBON
CATABOLITE
REPRESSION
IN
THE
GLUCOSE
KINASE
GLKA
.
6
THE
PATHWAY-SPECIFIC
REGULATORS
MALR
AND
GYLR
.
6
THE
PLEITROPIC
PROTEIN
BLDB
.
7
THE
CRRAI
GENE
LOCUS
.
9
MODEL
OF
CARBON
REGULATION
IN
5.
COELICOLOR
.
9
2.4
SCOPE
OF
THE
THESIS
.
.
.
3
MATERIALS
AND
METHODS
.
11
3.1
MATERIALS
.
.
.
.
.
11
3.1.1
CHEMICALS,
EQUIPMENT,
AND
OLIGONUCLEOTIDES
.
11
3.1.2
BACTERIAL
STRAINS
AND
PLASMIDS
.
.
.
15
3
J
MEDIA,
BUFFERS,
AND
SOLUTIONS
14
3.2.1
MEDIA
.
16
3.2.2
BUFFERS
AND
SOLUTIONS
.
17
33
METHODS
18
3.3.1
GENERAL
METHODS
.
18
3.3.2
GROWTH
OF
BACTERIA
.
18
3.3.3
TRANSFORMATION
OF
E.
COLI
.
19
3.3
4
TRANSFORMATION
OF
S.
COELICOLOR
.
19
3.4
METHODS
FOR
PROTEIN
PURIFICATION
AND
CHARACTERIZATION.
_
.
20
3.4
1
DENATURING
POLYACRYLAMIDE
GEL
ELECTROPHORESIS
(PAGE)
.
20
3.4.2
NATIVE
PAGE
.
20
3.43
SILVER-STAINING
OF
PROTEINS
.
20
3
4.4
SONIFICATION
.
21
3.4.5
DETERMINATION
OF
PROTEIN
CONCENTRATIONS
.
21
3.4.6
PURIFICATION
OF
HIS
6
-TAGGED-GLKA,
-GYLR,
-BLDB,
AND
-MALR
A
DBD
.
21
3.4.7
PURIFICATION
OF
NATIVE
BLDB
.
21
3.4.8
PURIFICATION
OF
GLKA
BINDING
PROTEINS
FROM
S',
COELICOLOR
EXTRACTS
.
21
3.4.9
SIZE
EXLUSION
CHROMATOGRAPHY
.
21
3.4.10
IN
VITRO
PHOSPHORYLATION
OF
PROTEINS
.
22
3.4.
1
1
IMMUNOLOGICAL
METHODS
.
22
3.4.12
SPECTROPHOTOMETRICAL
METHODS
.
23
3.4.13
SURFACE
PLASMON
RESONANCE
(SPR)
.23
3.4.14
DETERMINATION
OF
GFP
FLUORESCENCE
.23
33
METHODS
FOR
NUCLEIC ACID
PURIFICATION
AND
MODIFICATION
24
3.5.1
PURIFICATION
OF
PLASMID
DNA
WITH
MODIFIED
ALKALINE
/
SDS
LYSIS
.
24
3.5.2
ISOLATION
OF
CHROMOSOMAL
DNA
.
24
3.5.3
DNA
GEL
ELECTROPHORESIS
.
24
3.5.4
ELUTION
OF
DNA
FRAGMENTS
.
24
3.5.5
DNA
SEQUENCING
.
24
3.5.6
DNA
MODIFYING
ENZYMES
.
25
3.5.7
POLYMERASE
CHAIN
REACTION
(PCR)
.
25
3.5
8
REVERSE
TRANSCRIPTION
PCR
(RT-PCR)
.
25
INDEX
II
3.5.9
SITE-DIRECTED
MUTAGENESIS
.
26
3.5.10
ELECTROPHORETIC
MOBILITY
SHIFT
ASSAY
(EMSA)
.
26
3.6
ENZYME
ASSAYS
27
3.6.1
^-GALACTOSIDASE
ASSAY
.
27
3.6.2
T
RANSPORT
ASSAY
.
27
3.6.3
GLUCOSE
KINASE
ASSAY
.
28
3.7
COMPUTER
ANALYSES.
_
_
.^.
M
.YY.
M
.^.^.^^.W.^.28
4
RESULTS
.
29
4.1
SEARCH
FOR
REGULATORY
CAPACITIES
OF
GLUCOSE
KINASE
IN
CCR
.
.29
4.1.1
EXPRESSION
LEVELS
OF
GLUCOSE
KINASE
IN
S
COELICOLOR
.
29
4.1
2
CONSTRUCTION
OF
A
GFP-BASED
REPORTER
SYSTEM
TO
MONITOR
GENE
EXPRESSION
.
30
4.1.3
CLONING
OF
GLKA
OF
STREPTOMYCES
HVTDANS
.32
4.1.4
OVERPRODUCTION
AND
PURIFICATION
OF
GLKA
.33
STABILITY
OF
GLKA
UNDER
DIFFERENT
STORAGE
CONDITIONS
.34
4.1.5
ANALYSIS
OF
GLKA
STRUCTURE
.35
CIRCULAR
DICHROISM
SPECTROSCOPY
AND
SECONDARY
STRUCTURE
PREDICTION
.35
SIZE
EXCLUSION
CHROMATOGRAPHY.
36
CRYSTALLIZATION
OF
GLKA
.
36
4.1.6
GLKA
INTERACTION
ANALYSES
.
37
IDENTIFICATION
OF
A
GLKA
BINDING ACTIVITY
IN
S.
COELICOLOR
.
37
PURIFICATION
OF
GLKA
BINDING
PROTEINS
FROM
5.
COELICOLOR
EXTRACTS
.
38
BLDB
.
39
MALR
.43
GYLR
.
45
TWO-HYBRID
ANALYSIS
.
47
4.1.7
ATP
AND
PEP-DEPENDENT
PHOSPHORYLATION
OF
GLKA
.
49
4.2
ANALYSIS
OF
GLKA
MUTANTS
.
.
.
.^.^.^.-.W.W-^.^31
4.2.1
MULTIPLE
SEQUENCE
ALIGNMENT
OF
ROK
FAMILY
PROTEINS
.
51
42.2
CONSTRUCTION
AND
PURIFICATION
OF
GLKA
MUTANTS
.
52
4.2.3
ONE-HYBRID
ANALYSIS
.53
4.2.4
ATP-BINDING
TO
GLKA
.55
43
GLUCOSE
TRANSPORT
IN
5.
COELICOLOR
.
^.^.YY^.56
4.3.1
GLUCOSE
UPTAKE
IN
WILD-TYPE
AND
AGLKA
CELLS
.
56
4.3.2
SEARCH
FOR
A
GLUCOSE
TRANSPORT
PROTEIN
BY
IN
SILICA
ANALYSIS
.58
4.3.3
TRANSMEMBRANE
MODEL
OF
GLCP
.
59
4.3.4
IDENTIFICATION
OF
THE
GLUCOSE
TRANSPORTER
GLCP
BY
COMPLEMENTATION
IN
E.
COLI
.
59
4.3.5
CLONING
OF
GLCP
OF
STREPTOMYCES
LIVIDANS
.
60
4.3.6
BIOCHEMICAL
CHARACTERIZATION
OF
GLCP
.
61
4.3.7
GENETIC
ORGANIZATION
OF
GLCP
GENES
.63
4.3.8
ANALYSIS
OF
GLCP
EXPRESSION
IN
S.
COELICOLOR
.
64
4.3.9
THE
ROLE
OF
GLUCOSE
KINASE
IN
GLUCOSE
TRANSPORT
.67
5
DISCUSSION
.
69
5.1
ANALYSIS
OF
GLKA
EXPRESSION
.
W.^.^.W.W.
.
M
.
M
.
M
.
M
.69
52
GFP
AS
A
REPORTER
FOR
GENE
EXPRESSION
IN
SI
COELICOLOR
.
M
.W.
M
.
MM
.
M
.
M
.
MMM
.
MM
.73
53
SEARCH
FOR
REGULATORY
CAPACITIES
OF
GLKA
.
~.74
PURIFICATION
OF
WILD-TYPE GLKA
AND
STRUCTURAL
ANALYSES
.
74
INTERACTION
ANALYSES
.75
PHOSPHORYLATION
BY
PTS
PROTEINS
.
77
ACTIVITY
OF
PURIFIED
BLDB.
MALR,
AND
GYLR
.
77
INDEX
III
5.4
PURIFICATION
AND
ANALYSIS
OF
GLKA
MUTANTS.
.
_
.
------------
.79
5.5
THE
GLUCOSE
TRANSPORT
PROTEIN
GLCP.
-------
.
------------
.
----------
.80
5.6
CURRENT
WORKING
MODEL
AND
FUTURE
PERSPECTIVES
-----------------------------------------------------------------
83
6
LITERATURE
.
85
7
APPENDIX
.
95
7.1
ABBREVIATION
INDEX
_
-
---------------------------------------
95
7.2
PLASMID
MAPS.
-
.
-
.97 |
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| author_GND | (DE-588)123554179 |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV014219249 |
| illustrated | Illustrated |
| indexdate | 2024-08-19T00:10:09Z |
| institution | BVB |
| language | English German |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-009749366 |
| oclc_num | 50465019 |
| open_access_boolean | |
| owner | DE-29 DE-29T DE-12 DE-355 DE-BY-UBR DE-83 DE-188 |
| owner_facet | DE-29 DE-29T DE-12 DE-355 DE-BY-UBR DE-83 DE-188 |
| physical | III, 100 S. Ill., graph. Darst. |
| publishDate | 2001 |
| publishDateSearch | 2001 |
| publishDateSort | 2001 |
| record_format | marc |
| spelling | Mahr, Kerstin 1973- Verfasser (DE-588)123554179 aut Carbon metabolism in Streptomyces coelicolor a molecular view to glucose kinase and its role in carbon regulation and glucose transport von Kerstin Mahr 2001 III, 100 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Erlangen-Nürnberg, Univ., Diss., 2001. - Enth. Zsfassung in dt. Sprache Erlangen-Nürnberg, Univ., Diss., 2001 Hochschulschrift gtt Genregulation (DE-588)4122166-7 gnd rswk-swf Streptomyces coelicolor (DE-588)4183669-8 gnd rswk-swf Kohlenstoffstoffwechsel (DE-588)4451425-6 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Streptomyces coelicolor (DE-588)4183669-8 s Kohlenstoffstoffwechsel (DE-588)4451425-6 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=009749366&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Mahr, Kerstin 1973- Carbon metabolism in Streptomyces coelicolor a molecular view to glucose kinase and its role in carbon regulation and glucose transport Hochschulschrift gtt Genregulation (DE-588)4122166-7 gnd Streptomyces coelicolor (DE-588)4183669-8 gnd Kohlenstoffstoffwechsel (DE-588)4451425-6 gnd |
| subject_GND | (DE-588)4122166-7 (DE-588)4183669-8 (DE-588)4451425-6 (DE-588)4113937-9 |
| title | Carbon metabolism in Streptomyces coelicolor a molecular view to glucose kinase and its role in carbon regulation and glucose transport |
| title_auth | Carbon metabolism in Streptomyces coelicolor a molecular view to glucose kinase and its role in carbon regulation and glucose transport |
| title_exact_search | Carbon metabolism in Streptomyces coelicolor a molecular view to glucose kinase and its role in carbon regulation and glucose transport |
| title_full | Carbon metabolism in Streptomyces coelicolor a molecular view to glucose kinase and its role in carbon regulation and glucose transport von Kerstin Mahr |
| title_fullStr | Carbon metabolism in Streptomyces coelicolor a molecular view to glucose kinase and its role in carbon regulation and glucose transport von Kerstin Mahr |
| title_full_unstemmed | Carbon metabolism in Streptomyces coelicolor a molecular view to glucose kinase and its role in carbon regulation and glucose transport von Kerstin Mahr |
| title_short | Carbon metabolism in Streptomyces coelicolor |
| title_sort | carbon metabolism in streptomyces coelicolor a molecular view to glucose kinase and its role in carbon regulation and glucose transport |
| title_sub | a molecular view to glucose kinase and its role in carbon regulation and glucose transport |
| topic | Hochschulschrift gtt Genregulation (DE-588)4122166-7 gnd Streptomyces coelicolor (DE-588)4183669-8 gnd Kohlenstoffstoffwechsel (DE-588)4451425-6 gnd |
| topic_facet | Hochschulschrift Genregulation Streptomyces coelicolor Kohlenstoffstoffwechsel |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009749366&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT mahrkerstin carbonmetabolisminstreptomycescoelicoloramolecularviewtoglucosekinaseanditsroleincarbonregulationandglucosetransport |