Pseudomonas taiwanensis VLB120 synthetic biology: parts, modules, and chassis:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Aachen
Apprimus Verlag
2023
|
| Ausgabe: | 1. Auflage |
| Schriftenreihe: | Applied Microbiology
Volume 31 |
| Schlagworte: | |
| Online-Zugang: | Volltext Inhaltsverzeichnis |
| Beschreibung: | XIII, 102 Seiten Illustrationen, Diagramme 21 cm x 14.8 cm, 194 g |
| ISBN: | 9783985551705 3985551707 |
| DOI: | 10.18154/RWTH-2023-08705 |
Internformat
MARC
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| 100 | 1 | |a Silva Neves, Dário |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Pseudomonas taiwanensis VLB120 synthetic biology: parts, modules, and chassis |c Dário Silva Neves |
| 250 | |a 1. Auflage | ||
| 264 | 1 | |a Aachen |b Apprimus Verlag |c 2023 | |
| 300 | |a XIII, 102 Seiten |b Illustrationen, Diagramme |c 21 cm x 14.8 cm, 194 g | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 490 | 1 | |a Applied Microbiology |v Volume 31 | |
| 502 | |b Dissertation |c RWTH Aachen University |d 2023 | ||
| 650 | 0 | 7 | |a Pseudomonas taiwanensis |0 (DE-588)1064240615 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Genexpression |0 (DE-588)4020136-3 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Biotechnologie |0 (DE-588)4069491-4 |2 gnd |9 rswk-swf |
| 653 | |a Mikrobiologie | ||
| 653 | |a Naturwissenschaft | ||
| 653 | |a Klimawandel | ||
| 653 | |a Biologie | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Pseudomonas taiwanensis |0 (DE-588)1064240615 |D s |
| 689 | 0 | 1 | |a Biotechnologie |0 (DE-588)4069491-4 |D s |
| 689 | 0 | 2 | |a Genexpression |0 (DE-588)4020136-3 |D s |
| 689 | 0 | |5 DE-604 | |
| 710 | 2 | |a Apprimus Verlag |0 (DE-588)1068101474 |4 pbl | |
| 776 | 0 | 8 | |i Erscheint auch als |n Online-Ausgabe, PDF |o https://doi.org/10.18154/RWTH-2023-08705 |
| 830 | 0 | |a Applied Microbiology |v Volume 31 |w (DE-604)BV042644516 |9 31 | |
| 856 | 4 | 1 | |u https://doi.org/10.18154/RWTH-2023-08705 |x Resolving-System |z kostenfrei |3 Volltext |
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| 999 | |a oai:aleph.bib-bvb.de:BVB01-034607390 | ||
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Datensatz im Suchindex
| _version_ | 1804185876840841216 |
|---|---|
| adam_text | TABLE
OF
CONTENTS
SUMMARY
...................................................................................................................................................
III
ZUSAMMENFASSUNG.....................................................................................................................................
V
LIST
OF
ABBREVIATIONS
................................................................................................................................
VII
LIST
OF
FIGURES
............................................................................................................................................
XI
LIST
OF
TABLES
...........................................................................................................................................
XIII
1.
GENERAL
INTRODUCTION
.......................................................................................................................
3
1.1.
TOWARDS
A
CIRCULAR
BIOECONOMY
BASED
ON
RENEWABLE
CARBON
.......................................................
3
1.1.1.
CLIMATE
CHANGE
AND
THE
URGE
FOR
A
CIRCULAR
BIOECONOMY
.......................................................
3
1.1.2.
INTEGRATION
OF
BIOECONOMY
AND
CIRCULAR
ECONOMY
THE
CONCEPT
OF
A
CIRCULAR
BIOECONOMY..
7
1.2.
CELL
FACTORIES
AND
THEIR
DEVELOPMENT
...............................................................................................
8
1.3.
PSEUDOMONAS
TAIWANENSIS
VLB120
AS
A
CELL
FACTORY
.....................................................................
11
1.4.
SCOPE
AND
OUTLINE
OF
THE
THESIS
......................................................................................................
13
2.
MATERIAL
AND
METHODS
....................................................................................................................
17
2.1.
MEDIA
AND
CULTURE
CONDITIONS
........................................................................................................
17
2.2.
PLASMID
AND
STRAIN
CONSTRUCTION
....................................................................................................
18
2.3.
GRAVIMETRIC
CELL
DRY
WEIGHT
DETERMINATION
...................................................................................
21
2.4.
GROWTH
RATE
DETERMINATION
............................................................................................................
21
2.5.
FLUORESCENT
MEASUREMENTS
...........................................................................................................
21
2.6.
SEQUENCE
STATISTICS
.........................................................................................................................
22
2.7.
FLOW
CYTOMETRY
AND
CELL
SORTING
....................................................................................................
22
2.8.
QPCR
ASSAYS
FOR
MRNA
STABILITY
ASSESSMENT
.................................................................................
23
2.9.
ANALYTICAL
METHODS
.........................................................................................................................
24
2.10.
LC-MS
ANALYSIS
OF
PROPIONYL-COA
...................................................................................................
24
2.11.
FERMENTATION
.................................................................................................................................
24
3.
RESULTS............................................................................................................................................
29
3.1.
CROSS-SPECIES
SYNTHETIC
PROMOTER
LIBRARY:
FINDING
COMMON
GROUND
BETWEEN
PSEUDOMONAS
TAIWANENSIS
VLB120
AND
ESCHERICHIA
COLI
................................................................................................
29
3.1.1.
ABSTRACT
................................................................................................................................
29
3.1.2.
INTRODUCTION
..........................................................................................................................
29
3.1.3.
RESULTS
AND
DISCUSSION
.........................................................................................................
30
3.1.3.1.
DESIGN
AND
GENERATION
OF
THE
SYNTHETIC
PROMOTER
LIBRARIES
......................................................
30
3.1.3.2.
SEQUENCE
DIVERSITY
OF
GENERATED
PROMOTER
..............................................................................
33
3.1.3.3.
CHARACTERIZATION
OF
THE
SYNTHETIC
PROMOTER
LIBRARIES
IN
E.
COLI
TOP10
.......................................
34
3.1.3.4.
CHARACTERIZATION
OF
THE
SYNTHETIC
PROMOTER
LIBRARIES
IN
PSEUDMONAS
TAIWANENSIS
VLB120
......
36
3.1.3.5.
COMMON
GROUND
AND
DISPARITIES
OF
07-DEPENDENT
PROMOTERS
BETWEEN
P.
TAIWANENSIS
VLB120
AND
E.
COLI
..............................................................................................................................................
38
TABLE
OF
CONTENTS
3.1.4.
CONCLUSION
..............................................................................................................................41
3.2.
PSEUDOMONAS
MRNA
2.0:
BOOSTING
GENE
EXPRESSION
THROUGH
ENHANCED
MRNA
STABILITY
AND
TRANSLATIONAL
EFFICIENCY
.............................................................................................................................
45
3.2.1.
ABSTRACT
...................................................................................................................................
45
3.2.2.
INTRODUCTION
.............................................................................................................................
45
3.2.3.
RESULTS
AND
DISCUSSION
............................................................................................................
47
3.2.3.1.
CHARACTERIZATION
OF
PLASMID-BASED,
CONSTITUTIVE
FLUORESCENT
PROTEIN
EXPRESSION......................
47
3.2.3.2.
CHARACTERIZATION
OF
PLASMID-BASED,
INDUCIBLE
FLUORESCENT
PROTEIN
EXPRESSION
...........................
50
3.2.3.3.
CHARACTERIZATION
OF
INDUCIBLE
FLUORESCENCE
EXPRESSION
OF
SINGLE,
GENOME-INTEGRATED
CONSTRUCTS51
3.2.3.4.
EVALUATION
OF
A
RECOMBINANT
ACETOIN
PRODUCTION
PATHWAY
EMPLOYING
THE
OPTIMIZED
EXPRESSION
CASSETTE
.................................................................................................................................................
52
3.2.3.5.
QPCR
BASED
ELUCIDATION
OF
MRNA
STABILITY
.................................................................................
53
3.2.4.
CONCLUSION
..............................................................................................................................
54
3.3.
EXPANDING
PSEUDOMONAS
TAIWANENSIS
VLB120
7
S
ACYL-COA
PORTFOLIO:
PROPIONATE
PRODUCTION
IN
MINERAL
SALT
MEDIUM
.................................................................................................................................
57
3.3.1.
ABSTRACT
...................................................................................................................................
57
3.3.2.
INTRODUCTION
.............................................................................................................................
57
3.3.3.
RESULTS
AND
DISCUSSION
.............................................................................................................
59
3.3.3.1.
EXTENDING
THE
ACYL-COA
PORTFOLIO
OF
P.
TAIWANENSIS
VLB120
BY
EXPRESSING
THE
SLEEPING
BEAUTY
MUTASE
..................................................................................................................................................
59
3.3.3.2.
PROPIONATE
PRODUCTION
IN
P.
TAIWANENSIS
VLB120
.....................................................................
60
3.3.3.3.
BENCHMARKING
OF
FED-BATCH
FERMENTATION
STRATEGIES
FOR
PROPIONATE
PRODUCTION
.......................
62
3.3.4.
CONCLUSION
..............................................................................................................................
65
4.
GENERAL
DISCUSSION
..........................................................................................................................
69
4.1.
DEVELOPMENT
OF
CROSS-SPECIES
GENETIC
TOOLS
..................................................................................
69
4.2.
DEVELOPMENT
OF
ENHANCED
GENE
EXPRESSION
ARCHITECTURES
............................................................
71
4.3.
EXTENDING
THE
ACYL-COA
PORTFOLIO
OF
PSEUDOMONAS
TAIWANENSIS
....................................................
71
4.4.
CONCLUSION
.......................................................................................................................................
73
APPENDIX
...................................................................................................................................................
75
REFERENCES
................................................................................................................................................
83
CURRICULUM
VITAE
.......................................................................................................................................
99
|
| adam_txt |
TABLE
OF
CONTENTS
SUMMARY
.
III
ZUSAMMENFASSUNG.
V
LIST
OF
ABBREVIATIONS
.
VII
LIST
OF
FIGURES
.
XI
LIST
OF
TABLES
.
XIII
1.
GENERAL
INTRODUCTION
.
3
1.1.
TOWARDS
A
CIRCULAR
BIOECONOMY
BASED
ON
RENEWABLE
CARBON
.
3
1.1.1.
CLIMATE
CHANGE
AND
THE
URGE
FOR
A
CIRCULAR
BIOECONOMY
.
3
1.1.2.
INTEGRATION
OF
BIOECONOMY
AND
CIRCULAR
ECONOMY
THE
CONCEPT
OF
A
CIRCULAR
BIOECONOMY.
7
1.2.
CELL
FACTORIES
AND
THEIR
DEVELOPMENT
.
8
1.3.
PSEUDOMONAS
TAIWANENSIS
VLB120
AS
A
CELL
FACTORY
.
11
1.4.
SCOPE
AND
OUTLINE
OF
THE
THESIS
.
13
2.
MATERIAL
AND
METHODS
.
17
2.1.
MEDIA
AND
CULTURE
CONDITIONS
.
17
2.2.
PLASMID
AND
STRAIN
CONSTRUCTION
.
18
2.3.
GRAVIMETRIC
CELL
DRY
WEIGHT
DETERMINATION
.
21
2.4.
GROWTH
RATE
DETERMINATION
.
21
2.5.
FLUORESCENT
MEASUREMENTS
.
21
2.6.
SEQUENCE
STATISTICS
.
22
2.7.
FLOW
CYTOMETRY
AND
CELL
SORTING
.
22
2.8.
QPCR
ASSAYS
FOR
MRNA
STABILITY
ASSESSMENT
.
23
2.9.
ANALYTICAL
METHODS
.
24
2.10.
LC-MS
ANALYSIS
OF
PROPIONYL-COA
.
24
2.11.
FERMENTATION
.
24
3.
RESULTS.
29
3.1.
CROSS-SPECIES
SYNTHETIC
PROMOTER
LIBRARY:
FINDING
COMMON
GROUND
BETWEEN
PSEUDOMONAS
TAIWANENSIS
VLB120
AND
ESCHERICHIA
COLI
.
29
3.1.1.
ABSTRACT
.
29
3.1.2.
INTRODUCTION
.
29
3.1.3.
RESULTS
AND
DISCUSSION
.
30
3.1.3.1.
DESIGN
AND
GENERATION
OF
THE
SYNTHETIC
PROMOTER
LIBRARIES
.
30
3.1.3.2.
SEQUENCE
DIVERSITY
OF
GENERATED
PROMOTER
.
33
3.1.3.3.
CHARACTERIZATION
OF
THE
SYNTHETIC
PROMOTER
LIBRARIES
IN
E.
COLI
TOP10
.
34
3.1.3.4.
CHARACTERIZATION
OF
THE
SYNTHETIC
PROMOTER
LIBRARIES
IN
PSEUDMONAS
TAIWANENSIS
VLB120
.
36
3.1.3.5.
COMMON
GROUND
AND
DISPARITIES
OF
07-DEPENDENT
PROMOTERS
BETWEEN
P.
TAIWANENSIS
VLB120
AND
E.
COLI
.
38
TABLE
OF
CONTENTS
3.1.4.
CONCLUSION
.41
3.2.
PSEUDOMONAS
MRNA
2.0:
BOOSTING
GENE
EXPRESSION
THROUGH
ENHANCED
MRNA
STABILITY
AND
TRANSLATIONAL
EFFICIENCY
.
45
3.2.1.
ABSTRACT
.
45
3.2.2.
INTRODUCTION
.
45
3.2.3.
RESULTS
AND
DISCUSSION
.
47
3.2.3.1.
CHARACTERIZATION
OF
PLASMID-BASED,
CONSTITUTIVE
FLUORESCENT
PROTEIN
EXPRESSION.
47
3.2.3.2.
CHARACTERIZATION
OF
PLASMID-BASED,
INDUCIBLE
FLUORESCENT
PROTEIN
EXPRESSION
.
50
3.2.3.3.
CHARACTERIZATION
OF
INDUCIBLE
FLUORESCENCE
EXPRESSION
OF
SINGLE,
GENOME-INTEGRATED
CONSTRUCTS51
3.2.3.4.
EVALUATION
OF
A
RECOMBINANT
ACETOIN
PRODUCTION
PATHWAY
EMPLOYING
THE
OPTIMIZED
EXPRESSION
CASSETTE
.
52
3.2.3.5.
QPCR
BASED
ELUCIDATION
OF
MRNA
STABILITY
.
53
3.2.4.
CONCLUSION
.
54
3.3.
EXPANDING
PSEUDOMONAS
TAIWANENSIS
VLB120
7
S
ACYL-COA
PORTFOLIO:
PROPIONATE
PRODUCTION
IN
MINERAL
SALT
MEDIUM
.
57
3.3.1.
ABSTRACT
.
57
3.3.2.
INTRODUCTION
.
57
3.3.3.
RESULTS
AND
DISCUSSION
.
59
3.3.3.1.
EXTENDING
THE
ACYL-COA
PORTFOLIO
OF
P.
TAIWANENSIS
VLB120
BY
EXPRESSING
THE
SLEEPING
BEAUTY
MUTASE
.
59
3.3.3.2.
PROPIONATE
PRODUCTION
IN
P.
TAIWANENSIS
VLB120
.
60
3.3.3.3.
BENCHMARKING
OF
FED-BATCH
FERMENTATION
STRATEGIES
FOR
PROPIONATE
PRODUCTION
.
62
3.3.4.
CONCLUSION
.
65
4.
GENERAL
DISCUSSION
.
69
4.1.
DEVELOPMENT
OF
CROSS-SPECIES
GENETIC
TOOLS
.
69
4.2.
DEVELOPMENT
OF
ENHANCED
GENE
EXPRESSION
ARCHITECTURES
.
71
4.3.
EXTENDING
THE
ACYL-COA
PORTFOLIO
OF
PSEUDOMONAS
TAIWANENSIS
.
71
4.4.
CONCLUSION
.
73
APPENDIX
.
75
REFERENCES
.
83
CURRICULUM
VITAE
.
99 |
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| any_adam_object_boolean | 1 |
| author | Silva Neves, Dário |
| author_facet | Silva Neves, Dário |
| author_role | aut |
| author_sort | Silva Neves, Dário |
| author_variant | n d s nd nds |
| building | Verbundindex |
| bvnumber | BV049346922 |
| collection | ebook |
| ctrlnum | (OCoLC)1401215089 (DE-599)DNB1300364777 |
| doi_str_mv | 10.18154/RWTH-2023-08705 |
| edition | 1. Auflage |
| format | Thesis Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV049346922 |
| illustrated | Illustrated |
| index_date | 2024-07-03T22:48:33Z |
| indexdate | 2024-07-10T10:02:12Z |
| institution | BVB |
| institution_GND | (DE-588)1068101474 |
| isbn | 9783985551705 3985551707 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-034607390 |
| oclc_num | 1401215089 |
| open_access_boolean | 1 |
| owner | DE-29T |
| owner_facet | DE-29T |
| physical | XIII, 102 Seiten Illustrationen, Diagramme 21 cm x 14.8 cm, 194 g |
| psigel | ebook |
| publishDate | 2023 |
| publishDateSearch | 2023 |
| publishDateSort | 2023 |
| publisher | Apprimus Verlag |
| record_format | marc |
| series | Applied Microbiology |
| series2 | Applied Microbiology |
| spelling | Silva Neves, Dário Verfasser aut Pseudomonas taiwanensis VLB120 synthetic biology: parts, modules, and chassis Dário Silva Neves 1. Auflage Aachen Apprimus Verlag 2023 XIII, 102 Seiten Illustrationen, Diagramme 21 cm x 14.8 cm, 194 g txt rdacontent n rdamedia nc rdacarrier Applied Microbiology Volume 31 Dissertation RWTH Aachen University 2023 Pseudomonas taiwanensis (DE-588)1064240615 gnd rswk-swf Genexpression (DE-588)4020136-3 gnd rswk-swf Biotechnologie (DE-588)4069491-4 gnd rswk-swf Mikrobiologie Naturwissenschaft Klimawandel Biologie (DE-588)4113937-9 Hochschulschrift gnd-content Pseudomonas taiwanensis (DE-588)1064240615 s Biotechnologie (DE-588)4069491-4 s Genexpression (DE-588)4020136-3 s DE-604 Apprimus Verlag (DE-588)1068101474 pbl Erscheint auch als Online-Ausgabe, PDF https://doi.org/10.18154/RWTH-2023-08705 Applied Microbiology Volume 31 (DE-604)BV042644516 31 https://doi.org/10.18154/RWTH-2023-08705 Resolving-System kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034607390&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p vlb 20230829 DE-101 https://d-nb.info/provenance/plan#vlb |
| spellingShingle | Silva Neves, Dário Pseudomonas taiwanensis VLB120 synthetic biology: parts, modules, and chassis Applied Microbiology Pseudomonas taiwanensis (DE-588)1064240615 gnd Genexpression (DE-588)4020136-3 gnd Biotechnologie (DE-588)4069491-4 gnd |
| subject_GND | (DE-588)1064240615 (DE-588)4020136-3 (DE-588)4069491-4 (DE-588)4113937-9 |
| title | Pseudomonas taiwanensis VLB120 synthetic biology: parts, modules, and chassis |
| title_auth | Pseudomonas taiwanensis VLB120 synthetic biology: parts, modules, and chassis |
| title_exact_search | Pseudomonas taiwanensis VLB120 synthetic biology: parts, modules, and chassis |
| title_exact_search_txtP | Pseudomonas taiwanensis VLB120 synthetic biology: parts, modules, and chassis |
| title_full | Pseudomonas taiwanensis VLB120 synthetic biology: parts, modules, and chassis Dário Silva Neves |
| title_fullStr | Pseudomonas taiwanensis VLB120 synthetic biology: parts, modules, and chassis Dário Silva Neves |
| title_full_unstemmed | Pseudomonas taiwanensis VLB120 synthetic biology: parts, modules, and chassis Dário Silva Neves |
| title_short | Pseudomonas taiwanensis VLB120 synthetic biology: parts, modules, and chassis |
| title_sort | pseudomonas taiwanensis vlb120 synthetic biology parts modules and chassis |
| topic | Pseudomonas taiwanensis (DE-588)1064240615 gnd Genexpression (DE-588)4020136-3 gnd Biotechnologie (DE-588)4069491-4 gnd |
| topic_facet | Pseudomonas taiwanensis Genexpression Biotechnologie Hochschulschrift |
| url | https://doi.org/10.18154/RWTH-2023-08705 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034607390&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV042644516 |
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