Transcriptional responses and transcriptional regulators of Gluconobacter oxydans 621H:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Jülich
Forschungszentrum Jülich
2012
|
| Schriftenreihe: | Schriften des Forschungszentrums Jülich
Reihe Gesundheit ; 51 |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XVI, 140 S. Ill., graph. Darst. |
| ISBN: | 9783893367801 |
Internformat
MARC
| LEADER | 00000nam a2200000 cb4500 | ||
|---|---|---|---|
| 001 | BV040149069 | ||
| 003 | DE-604 | ||
| 005 | 00000000000000.0 | ||
| 007 | t | ||
| 008 | 120523s2012 ad|| m||| 00||| eng d | ||
| 020 | |a 9783893367801 |9 978-3-89336-780-1 | ||
| 035 | |a (OCoLC)796237717 | ||
| 035 | |a (DE-599)HBZHT017198884 | ||
| 040 | |a DE-604 |b ger |e rakwb | ||
| 041 | 0 | |a eng | |
| 049 | |a DE-12 | ||
| 100 | 1 | |a Schweikert, Stefanie Karolin |e Verfasser |0 (DE-588)1022644726 |4 aut | |
| 245 | 1 | 0 | |a Transcriptional responses and transcriptional regulators of Gluconobacter oxydans 621H |c Stefanie Karolin Schweikert |
| 264 | 1 | |a Jülich |b Forschungszentrum Jülich |c 2012 | |
| 300 | |a XVI, 140 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 490 | 1 | |a Schriften des Forschungszentrums Jülich : Reihe Gesundheit |v 51 | |
| 502 | |a Zugl.: Düsseldorf, Univ., Diss., 2011 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 830 | 0 | |a Schriften des Forschungszentrums Jülich |v Reihe Gesundheit ; 51 |w (DE-604)BV035800060 |9 51 | |
| 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=025005821&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 999 | |a oai:aleph.bib-bvb.de:BVB01-025005821 | ||
Datensatz im Suchindex
| _version_ | 1804149172371193856 |
|---|---|
| adam_text | IMAGE 1
CONTENTS
PUBLICATIONS I L L
ABSTRACT V
ZUSAMMENFASSUNG V I I
LIST O F FIGURES X I I
LIST O F TABLES X I V
ABBREVIATIONS X V I
1 INTRODUCTION 1
1.1 GENERAL INFORMATION ABOUT G. OXYDANS 6 2 1 H 1
1.2 MANNITOL AND FRUCTOSE METABOLISM IN G. OXYDANS 3
1.3 IRON UPTAKE IN BACTERIA 5
1.4 TRANSCRIPTIONAL REGULATORS IN BACTERIA 5
1.5 T H E FUMARATE/NITRATE REDUCTION REGULATOR F N R 8
1.6 A I M S O F THIS THESIS 9
2 MATERIALS AND METHODS 11
2.1 GENERAL MATERIALS A N D PROCEDURES 11
2.1.1 GENERAL MATERIALS 11
2.1.2 GENERAL PROCEDURES 11
2.1.3 GENERAL-USE BUFFERS A N D M E D I A 12
2.2 STRAINS, PLASMIDS A N D OLIGONUCLEOTIDES 13
2.2.1 GLUCONOBACTER A N D E. COLI STRAINS 13
2.2.2 PLASMIDS 14
2.2.3 OLIGONUCLEOTIDES 15
2.3 I N SILICO ANALYSIS FOR DETERMINING THE TRANSCRIPTIONAL REGULATOR
REPERTOIRE . . . . 17
2.4 CLONING AND D N A TECHNIQUES 18
2.4.1 P C R 18
2.4.2 LIGATION 2 0
2.4.3 CLONING O F PLASMIDS 2 0
2.4.4 TRANSFORMATION O F MICROORGANISMS 2 2
2.4.5 MARKER-FREE DELETION O F GENES IN G. OXYDANS B Y THE S A C B M E T
H O D . . . . 2 3
2.5 CULTIVATION CONDITIONS AND H P L C ANALYTICS 2 4
HTTP://D-NB.INFO/1021825514
IMAGE 2
X CONTENTS
2.5.1 CULTIVATION IN SHAKING FLASKS 2 4
2.5.2 CULTIVATION IN A PARALLEL BIOREACTOR SYSTEM 2 4
2.5.3 H P L C ANALYTICS 2 5
2.6 R N A RELATED METHODS 2 5
2.6.1 SAMPLING A N D ISOLATION O F R N A 2 5
2.6.2 TRANSCRIPTIONAL ANALYSES B Y USING OLIGONUCLEOTIDE MICROARRAYS 2 6
2.6.3 DETERMINATION O F TRANSCRIPTIONAL START SITES B Y 5 R A C E 2 7
2.7 BIOCHEMICAL TECHNIQUES 2 8
2.7.1 S D S P A G E 2 8
2.7.2 PROTEIN EXPRESSION AND PURIFICATION O F G O X R 2 8
2.7.3 (3-GALACTOSIDASE ASSAY 3 0
2.7.4 ACTIVITY O F PENTOSE PHOSPHATE PATHWAY E N Z Y M E S 3 0
2.7.5 ELECTROPHORETIC MOBILITY SHIFT ASSAYS ( E M S A ) 3 3
3 RESULTS 3 4
3.1 TRANSCRIPTIONAL RESPONSE O F G. OXYDANS TO IRON LIMITATION 3 4
3.2 COMPARISON O F GLOBAL G E N E EXPRESSION FOR THE CARBON SOURCES
MANNITOL A N D FRUC
TOSE 3 6
3.2.1 GROWTH CHARACTERISTICS O F CULTURES USING MANNITOL O R FRUCTOSE AS
SUBSTRATES 3 6
3.2.2 TRANSCRIPTIONAL ANALYSIS O F FRUCTOSE-VS. MANNITOL-GROWN CELLS 3 9
3.2.3 TRANSCRIPTIONAL ANALYSIS O F 2 N D VS. 1ST GROWTH P H A S E USING
MANNITOL A S
CARBON SOURCE 4 2
3.2.4 E N Z Y M E ACTVITY ASSAYS O F SELECTED P P P PROTEINS 4 5
3.3 T H E REPERTOIRE O F TRANSCRIPTIONAL REGULATORS IN G. OXYDANS: A
BIOINFORMATIC ANALYSIS 4 6
3.3.1 C H A N G E IN EXPRESSION O F TRANSCRIPTIONAL REGULATORS 5 7
3.4 INITIAL CHARACTERIZATION O F SELECTED TRANSCRIPTIONAL REGULATORS:
GROWTH PROPERTIES
A N D GLOBAL G E N E EXPRESSION O F MUTANT STRAINS 5 8
3.4.1 GROWTH BEHAVIOUR O F CANDIDATE REGULATOR DELETION STRAINS 5 9
3.4.2 TRANSCRIPTIONAL ANALYSES O F REGULATOR MUTANTS 61
3.4.3 S U M M A R Y O F THE SCREENING EXPERIMENTS WITH REGULATOR
DELETION STRAINS . 6 5
3.5 CHARACTERISATION O F AN FNR-LIKE TRANSCRIPTIONAL REGULATOR
CONTAINING A PUTATIVE
IRON-SULFUR CLUSTER: EVIDENCE F O R INVOLVEMENT IN REDOX METABOLISM 6 6
3.5.1 T H E IN SILICO ANALYSIS O F G O X 0 9 7 4 SUGGESTS A N FNR-LIKE
FUNCTION . . . . 6 6
3.5.2 PUTATIVE REGULON O F G O X 0 9 7 4 6 7
3.5.3 PUTATIVE TARGET GENES O F G O X 0 9 7 4 7 0
3.5.4 O X Y G E N DEPENDENT ACTIVITY O F G O X 0 9 7 4 7 1
3.5.4.1 REACTION O N O X Y G E N LIMITATION 71
3.5.4.2 REACTION O N O X Y G E N EXCESS 7 2
3.5.5 BINDING M O T I F O F G Q X 0 9 7 4 7 3
IMAGE 3
CONTENTS X I
3.5.6 IDENTIFICATION O F TRANSCRIPTIONAL START SITES (TSS) O F CYDA A N
D P N T A L . . . 7 4
3.5.7 PURIFICATION O F G O X 0 9 7 4 7 5
3.5.8 OXYGEN-DEPENDENT SPECTRA O F G O X 0 9 7 4 7 6
3.5.9 ELECTROPHORETIC MOBILITY SHIFT ASSAYS ( E M S A S ) WITH G O X 0 9
7 4 7 7
4 DISCUSSION 79
4.1 IRON LIMITATION IN G. OXYDANS LEADS TO INCREASED M R N A LEVELS O F
GENES FOR IRON
UPTAKE SYSTEMS AND FLAGELLA SYNTHESIS 7 9
4.2 T H E SUBSTRATES MANNITOL A N D FRUCTOSE DIFFER IN PATHWAY
PREFERENCE F O R CARBON
ASSIMILATION 81
4.3 T H E DETERMINATION O F THE REPERTOIRE O F TRANSCRIPTIONAL
REGULATORS IN G. O X Y D A N S
REVEALED CANDIDATES FOR FURTHER STUDIES 8 8
4.3.1 T H E EXPRESSION LEVEL O F T R S IN GLOBAL MICROARRAY ANALYSES
REVEALED
PROMISING CANDIDATES 91
4.4 T H E INITIAL CHARACTERISATION O F T R DELETION MUTANTS REVEALED A
PROMISING CANDI
DATE ACTIVE IN REDOX A N D ENERGY METABOLISM 9 2
4.4.1 PUTATIVE FUNCTION O F HXLR-TYPE REGULATOR G O X 0 4 9 8 9 2
4.4.2 PUTATIVE FUNCTION O F LYSR-TYPE REGULATOR G O X 1 3 3 7 9 2
4.4.3 PUTATIVE FUNCTION O F TETR-TYPE REGULATOR G O X 0 5 6 3 9 3
4.4.4 PUTATIVE FUNCTION O F LYSR-TYPE REGULATOR G O X 1711 9 4
4.4.5 PUTATIVE FUNCTION O F GALR/LACI-LIKE REGULATOR G O X 0 5 7 7 9 4
4.4.6 PUTATIVE FUNCTION O F FNR-LIKE REGULATOR G O X 0 9 7 4 9 5
4.4.7 S U M M A R Y O F REGULATOR MUTANT PHENOTYPE STUDIES IN G. OXYDANS
9 5
4.5 G O X 0 9 7 4 IS A MASTER REGULATOR IN G. OXYDANS THAT IS INVOLVED
IN REDOX AND
ENERGY METABOLISM 9 6
4.5.1 G O X 0 9 7 4 DISPLAYS A REGULON O F GENES RELATED TO ENERGY A N D
REDOX
METABOLISM; THREE TARGET GENES WERE EXPERIMENTALLY PROVEN 9 6
4.5.2 T H E BINDING M O T I F O F G O X 0 9 7 4 AND PUTATIVE REGULATORY
MECHANISMS . . 9 8
4.5.3 T H E BIOCHEMICAL PROPERTIES O F G O X 0 9 7 4 INDICATE THE
INVOLVEMENT O F A N
IRON-SULFUR CLUSTER A N D O X Y G E N LABILITY 9 9
4.5.4 G O X 0 9 7 4 ACTIVITY AS TRANSCRIPTIONAL REGULATOR DEPENDS O N
OXYGEN . . . 100
4.5.5 S U M M A R Y O F G O X 0 9 7 4 FUNCTION 102
REFERENCES 103
APPENDIX 114
ACKNOWLEDGEMENTS 141
DECLARATION 142
|
| any_adam_object | 1 |
| author | Schweikert, Stefanie Karolin |
| author_GND | (DE-588)1022644726 |
| author_facet | Schweikert, Stefanie Karolin |
| author_role | aut |
| author_sort | Schweikert, Stefanie Karolin |
| author_variant | s k s sk sks |
| building | Verbundindex |
| bvnumber | BV040149069 |
| ctrlnum | (OCoLC)796237717 (DE-599)HBZHT017198884 |
| format | Thesis Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01376nam a2200313 cb4500</leader><controlfield tag="001">BV040149069</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">00000000000000.0</controlfield><controlfield tag="007">t</controlfield><controlfield tag="008">120523s2012 ad|| m||| 00||| eng d</controlfield><datafield tag="020" ind1=" " ind2=" "><subfield code="a">9783893367801</subfield><subfield code="9">978-3-89336-780-1</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)796237717</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)HBZHT017198884</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-12</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Schweikert, Stefanie Karolin</subfield><subfield code="e">Verfasser</subfield><subfield code="0">(DE-588)1022644726</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Transcriptional responses and transcriptional regulators of Gluconobacter oxydans 621H</subfield><subfield code="c">Stefanie Karolin Schweikert</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="a">Jülich</subfield><subfield code="b">Forschungszentrum Jülich</subfield><subfield code="c">2012</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">XVI, 140 S.</subfield><subfield code="b">Ill., graph. Darst.</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">n</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">nc</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="490" ind1="1" ind2=" "><subfield code="a">Schriften des Forschungszentrums Jülich : Reihe Gesundheit</subfield><subfield code="v">51</subfield></datafield><datafield tag="502" ind1=" " ind2=" "><subfield code="a">Zugl.: Düsseldorf, Univ., Diss., 2011</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4113937-9</subfield><subfield code="a">Hochschulschrift</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="830" ind1=" " ind2="0"><subfield code="a">Schriften des Forschungszentrums Jülich</subfield><subfield code="v">Reihe Gesundheit ; 51</subfield><subfield code="w">(DE-604)BV035800060</subfield><subfield code="9">51</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">DNB Datenaustausch</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025005821&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="999" ind1=" " ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-025005821</subfield></datafield></record></collection> |
| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV040149069 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T00:18:48Z |
| institution | BVB |
| isbn | 9783893367801 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-025005821 |
| oclc_num | 796237717 |
| open_access_boolean | |
| owner | DE-12 |
| owner_facet | DE-12 |
| physical | XVI, 140 S. Ill., graph. Darst. |
| publishDate | 2012 |
| publishDateSearch | 2012 |
| publishDateSort | 2012 |
| publisher | Forschungszentrum Jülich |
| record_format | marc |
| series | Schriften des Forschungszentrums Jülich |
| series2 | Schriften des Forschungszentrums Jülich : Reihe Gesundheit |
| spelling | Schweikert, Stefanie Karolin Verfasser (DE-588)1022644726 aut Transcriptional responses and transcriptional regulators of Gluconobacter oxydans 621H Stefanie Karolin Schweikert Jülich Forschungszentrum Jülich 2012 XVI, 140 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Schriften des Forschungszentrums Jülich : Reihe Gesundheit 51 Zugl.: Düsseldorf, Univ., Diss., 2011 (DE-588)4113937-9 Hochschulschrift gnd-content Schriften des Forschungszentrums Jülich Reihe Gesundheit ; 51 (DE-604)BV035800060 51 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025005821&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schweikert, Stefanie Karolin Transcriptional responses and transcriptional regulators of Gluconobacter oxydans 621H Schriften des Forschungszentrums Jülich |
| subject_GND | (DE-588)4113937-9 |
| title | Transcriptional responses and transcriptional regulators of Gluconobacter oxydans 621H |
| title_auth | Transcriptional responses and transcriptional regulators of Gluconobacter oxydans 621H |
| title_exact_search | Transcriptional responses and transcriptional regulators of Gluconobacter oxydans 621H |
| title_full | Transcriptional responses and transcriptional regulators of Gluconobacter oxydans 621H Stefanie Karolin Schweikert |
| title_fullStr | Transcriptional responses and transcriptional regulators of Gluconobacter oxydans 621H Stefanie Karolin Schweikert |
| title_full_unstemmed | Transcriptional responses and transcriptional regulators of Gluconobacter oxydans 621H Stefanie Karolin Schweikert |
| title_short | Transcriptional responses and transcriptional regulators of Gluconobacter oxydans 621H |
| title_sort | transcriptional responses and transcriptional regulators of gluconobacter oxydans 621h |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025005821&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV035800060 |
| work_keys_str_mv | AT schweikertstefaniekarolin transcriptionalresponsesandtranscriptionalregulatorsofgluconobacteroxydans621h |