Real time analysis of competence development in Bacillus subtilis:
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| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
2012
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | III, 129 S. Ill., graph. Darst. |
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MARC
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| 003 | DE-604 | ||
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| 100 | 1 | |a Karlshofer, Alexander |d 1981- |e Verfasser |0 (DE-588)1025123700 |4 aut | |
| 245 | 1 | 0 | |a Real time analysis of competence development in Bacillus subtilis |c vorgelegt von Alexander Karlshofer |
| 246 | 1 | |a Real-time analysis of competence development in Bacillus subtilis | |
| 264 | 1 | |c 2012 | |
| 300 | |a III, 129 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a Münster (Westfalen), Univ., Diss., 2012 | ||
| 650 | 0 | 7 | |a Kompetenz |g Bakteriologie |0 (DE-588)4309193-3 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Heubacillus |0 (DE-588)4242465-3 |2 gnd |9 rswk-swf |
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Record in the Search Index
| _version_ | 1804149426009145344 |
|---|---|
| adam_text | IMAGE 1
CONTENTS
CONTENTS I
1 INTRODUCTION 1
1.1 QUORUM SENSING AND CELLULAR DIFFERENTIATION 1
1.2 REGULATION NETWORK O F C O M K 4
1.2.1 TRANSCRIPTIONAL REGULATION O F C O M K 5
1.2.2 POST-TRANSLATIONAL REGULATION O F C O M K 6
1.3 BIMODALITY O F C O M K EXPRESSION 7
1.4 ROLE O F NOISE IN GENE EXPRESSION 8
1.5 MATHEMATICAL MODELING O F COMPETENCE DEVELOPMENT . 10
1.6 TRANSFORMATION 15
1.6.1 THE D N A UPTAKE MACHINERY 15
1.6.2 D N A BINDING 16
1.6.3 PASSING THE CELL ENVELOPE 17
1.6.4 RECOMBINATION AND DOUBLE STRAND BREAK REPAIR 18
1.6.5 RECN 19
2 OBJECTIVES O F THIS THESIS 21
3 MATERIALS AND METHODS 23
3.1 BIOLOGY 23
3.1.1 LIST O F STRAINS 23
3.1.2 CULTIVATING BACTERIA 2 4
3.1.3 GROWING BACTERIA TO COMPETENCE 25
3.1.4 TRANSFORMATION O F BACTERIA 26
3.1.5 MOLECULAR BIOLOGY 27
3.1.5.1 ISOLATION O F D N A 27
3.1.5.2 PHENOL/CHLOROFORM-EXTRACTION 27
3.1.5.3 P C R 28
3.1.5.4 HYDROLYSIS O F D N A WITH THE HELP O F RESTRICTION
ENDONUCLEASES. 30
3.1.5.5 DEPHOSPHORYLATION O F VECTORS 30
3.1.5.6 LIGATION 30
3.1.6 GENETIC ENGINEERING 31
3.1.6.1 RELEVANT PLASMIDS 31
3.1.6.2 OLIGONUCLEOTIDES 31
3.1.6.3 CONSTRUCTION O F RECN-GFP 33
3.1.6.4 CONSTRUCTION O F P COM ^-CFP 33
I
HTTP://D-NB.INFO/1026496721
IMAGE 2
CONTENTS
3.1.6.5 CONSTRUCTION O F AOPP 33
3.1.6.6 CONSTRUCTION O F COMGA- CFP 34
3.2 PREPARATIVE METHODS 37
3.2.1 FLUORESCENCE MICROSCOPY 37
3.2.2 PREPARATION O F POLYSTYRENE COATED COVER SLIDES 37
3.2.3 PREPARATION O F AGAR COVERED BIOLOGICAL SAMPLES USING IB1DI WELLS
...38 3.2.4 IMAGE ACQUISITION WITH FLUORESCENCE MICROSCOPY 38
3.3 IMAGE ANALYSIS 39
3.3.1 IMAGE PROCESSING 39
3.3.2 EVALUATION O F THE SWITCHING KINETICS DATA 40
3.3.3 DETERMINATION O F THE CELL LENGTH 41
3.3.4 STATISTICAL ANALYSIS - PEARSON CORRELATION COEFFICIENT 41
3.3.5 THE TRACKING ALGORITHM 43
3.3.6 MEAN SQUARED DISPLACEMENT 43
3.3.7 SIMULATED DIFFUSION - A SYNTHETIC APPROACH 46
3.3.8 FLUORESCENT DYE-MATURATION TIME 47
RESULTS 49
4.1 QUANTITATIVE RELATION BETWEEN C O M K AND LATE COMPETENCE PROTEINS 4
9
4.1.1 THE EXPRESSION O F LATE COMPETENCE GENES BEGINS 20 MINUTES AFTER
THE
ACTIVATION O F P CO, A- 4 9
4.1.2 EXPRESSION KINETICS O F C O M K AND O F LATE COMPETENCE GENES 53
4.1.3 BOTH THE EXPRESSION LEVELS AND SWITCHING KINETICS O F C O M K AND
LATE
COMPETENCE GENES ARE UNCORRECTED 55
4.1.4 COMPETENCE RULES CELL CYCLE OVER SEVERAL HOURS 60
4.1.5 GENETIC SWITCHING INTO COMPETENCE OCCURS PREFERENTIALLY IN THE
CENTER
O F THE CELL CYCLE 63
4.1.6 GROWTH RATE IMPAIRMENT IS INDEPENDENT O F BOTH THE MAXIMUM
EXPRESSION LEVEL AND THE SWITCHING KINETICS O F COMPETENCE PROTEINS
...64
4.2 ANALYSIS O F PUTATIVE COMPETENCE INHIBITORY SUBSTANCES 67
4.2.1 CONDITIONED GROWTH MEDIUM AFFECTS THE FRACTION O F COMPETENT CELLS
IN
AN OPP KNOCKOUT 68
4.2.2 THE FRACTION O F COMPETENT CELLS IS UNAFFECTED B Y SYNTHESIZED
CSF....69
4.3 CHARACTERIZATION O F RECN-FOCUS DYNAMICS 71
4.3.1 ALL RECN IS CONCENTRATED IN A SINGLE FOCUS PER CELL 71
4.3.2 DYNAMICS O F RECN FOCI 72
4.3.3 SPATIAL CONFINEMENT RESTRICTS FREE DIFFUSION 74
4.3.4 LOCALIZATION O F RECN FOCI 76
DISCUSSION 79
II
IMAGE 3
CONTENTS
5.1 QUANTITATIVE RELATION BETWEEN C O M K AND LATE COMPETENCE PROTEINS
IN B . SUBTILIS 79
5.1.1 EXPRESSION DYNAMICS O F C O M K AND LATE COMPETENCE GENES 79
5.1.2 CORRELATION ANALYSIS O F EXPRESSION PROPERTIES O F C O M K AND
LATE COMPETENCE GENES 80
5.2 PHYSIOLOGICAL CHANGES DURING COMPETENCE 84
5.2.1 THE COMPETENCE INDUCED GROWTH ARREST 84
5.2.2 TIME POINT O F SWITCHING DURING GROWTH PHASE 86
5.3 THE DECLINE O F COMPETENCE DEVELOPMENT 86
5.3.1 EFFECTS O F PURE SYNTHESIZED CSF ON B. SUBTILIS CULTURES GROWING T
O
COMPETENCE 87
5.3.2 EFFECTS O F CONDITIONED SUPERNATANT ON OPP MUTANT CULTURES GROWING
T O COMPETENCE 88
5.4 CHARACTERIZATION O F RECN-FOCI DYNAMICS 89
5.4.1 GENERAL ASPECTS O F RECN FOCI DYNAMICS 89
5.4.2 THE NATURE O F THE OBSERVED RECN-GFP-FOCI MOVEMENT 90
5.4.3 RECN-GFP-FOCI PREFERENTIALLY LOCATE NEAR THE CELL POLES 93
6 OUTLOOK 95
7 SUMMARY 97
8 ZUSAMMENFASSUNG 99
9 APPENDIX 103
A. PLASMID MAPS 103
ACRONYMS 108
LIST O F FIGURES 110
LIST O F TABLES 112
LITERATURE 113
WEBLINKS 126
LEBENSLAUF 127
DANKSAGUNG 129
III
|
| any_adam_object | 1 |
| author | Karlshofer, Alexander 1981- |
| author_GND | (DE-588)1025123700 |
| author_facet | Karlshofer, Alexander 1981- |
| author_role | aut |
| author_sort | Karlshofer, Alexander 1981- |
| author_variant | a k ak |
| building | Verbundindex |
| bvnumber | BV040383124 |
| ctrlnum | (OCoLC)815888245 (DE-599)HBZHT017353646 |
| dewey-full | 579.36217 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 579 - Microorganisms, fungi & algae |
| dewey-raw | 579.36217 |
| dewey-search | 579.36217 |
| dewey-sort | 3579.36217 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| format | Thesis Book |
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| genre_facet | Hochschulschrift |
| id | DE-604.BV040383124 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T00:22:50Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-025236607 |
| oclc_num | 815888245 |
| open_access_boolean | |
| owner | DE-188 DE-355 DE-BY-UBR DE-29T |
| owner_facet | DE-188 DE-355 DE-BY-UBR DE-29T |
| physical | III, 129 S. Ill., graph. Darst. |
| publishDate | 2012 |
| publishDateSearch | 2012 |
| publishDateSort | 2012 |
| record_format | marc |
| spelling | Karlshofer, Alexander 1981- Verfasser (DE-588)1025123700 aut Real time analysis of competence development in Bacillus subtilis vorgelegt von Alexander Karlshofer Real-time analysis of competence development in Bacillus subtilis 2012 III, 129 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Münster (Westfalen), Univ., Diss., 2012 Kompetenz Bakteriologie (DE-588)4309193-3 gnd rswk-swf Heubacillus (DE-588)4242465-3 gnd rswk-swf Genexpression (DE-588)4020136-3 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Heubacillus (DE-588)4242465-3 s Kompetenz Bakteriologie (DE-588)4309193-3 s Genexpression (DE-588)4020136-3 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025236607&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Karlshofer, Alexander 1981- Real time analysis of competence development in Bacillus subtilis Kompetenz Bakteriologie (DE-588)4309193-3 gnd Heubacillus (DE-588)4242465-3 gnd Genexpression (DE-588)4020136-3 gnd |
| subject_GND | (DE-588)4309193-3 (DE-588)4242465-3 (DE-588)4020136-3 (DE-588)4113937-9 |
| title | Real time analysis of competence development in Bacillus subtilis |
| title_alt | Real-time analysis of competence development in Bacillus subtilis |
| title_auth | Real time analysis of competence development in Bacillus subtilis |
| title_exact_search | Real time analysis of competence development in Bacillus subtilis |
| title_full | Real time analysis of competence development in Bacillus subtilis vorgelegt von Alexander Karlshofer |
| title_fullStr | Real time analysis of competence development in Bacillus subtilis vorgelegt von Alexander Karlshofer |
| title_full_unstemmed | Real time analysis of competence development in Bacillus subtilis vorgelegt von Alexander Karlshofer |
| title_short | Real time analysis of competence development in Bacillus subtilis |
| title_sort | real time analysis of competence development in bacillus subtilis |
| topic | Kompetenz Bakteriologie (DE-588)4309193-3 gnd Heubacillus (DE-588)4242465-3 gnd Genexpression (DE-588)4020136-3 gnd |
| topic_facet | Kompetenz Bakteriologie Heubacillus Genexpression Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025236607&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT karlshoferalexander realtimeanalysisofcompetencedevelopmentinbacillussubtilis |