Regulation of virulence-associated traits of the human fungal pathogen Candida albicans by nitrogen availability:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | German |
| Veröffentlicht: |
2013
|
| Schlagworte: | |
| Online-Zugang: | Volltext Inhaltsverzeichnis |
| Beschreibung: | Zsfassung in dt. und engl. Sprache |
| Beschreibung: | IV, 158, IV S. Ill., graph. Darst. |
Internformat
MARC
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| 100 | 1 | |a Dunkel, Nico |d 1983- |e Verfasser |0 (DE-588)104340466X |4 aut | |
| 245 | 1 | 0 | |a Regulation of virulence-associated traits of the human fungal pathogen Candida albicans by nitrogen availability |c von Nico Dunkel |
| 246 | 1 | 3 | |a Regulation Virulenz-assoziierter Faktoren im humanpathogenen Pilz Candida albicans durch Stickstoffverfügbarkeit |
| 264 | 1 | |c 2013 | |
| 300 | |a IV, 158, IV S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Zsfassung in dt. und engl. Sprache | ||
| 502 | |a Würzburg, Univ., Diss., 2013 | ||
| 650 | 0 | 7 | |a Candida albicans |0 (DE-588)4147244-5 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Proteasen |0 (DE-588)4047521-9 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Stickstoff |0 (DE-588)4125639-6 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Regulation |0 (DE-588)4049075-0 |2 gnd |9 rswk-swf |
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| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Candida albicans |0 (DE-588)4147244-5 |D s |
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| 912 | |a ebook | ||
| 999 | |a oai:aleph.bib-bvb.de:BVB01-026829547 | ||
Datensatz im Suchindex
| _version_ | 1804151477672869888 |
|---|---|
| adam_text | CONTENT
SUMMARY
ZUSAMMENFASSUNG
1 INTRODUCTION 1
1.1 CANDIDA ALBICANS 2
1.2 GENETICS 3
1.3 DRUG RESISTANCE 5
1.4 ADHERENCE 6
1.5 BIOFILM FORMATION 6
1.6 MORPHOGENESIS 7
1.7 SECRETED HYDROLYTIC ENZYMES 11
1.8 STRESS RESPONSE 16
1.9 METABOLIE ADAPTION 18
1.10 NITROGEN REGULATION 19
1.11 AIM OFTHIS STUDY 28
2 RESULTS 29
2.1 STPL, A TRANSCRIPTIONAL REGULATOR REQUIRED FOR GROWTH ON PROTEINS 29
2.1.1 CONSTRUCTION OF THE HA-TAGGED TRANSCRIPTIONAL REGULATOR STPL 29
2.1.2 HA-TAGGED STPL IS A FUNCTIONAL TRANSCRIPTION FACTOR 30
2.1.3 GENOME-WIDE BINDING OF STPL-HA TO TARGET SEQUENCES 31
2.2 SAP2, A MAJOR SECRETED ASPARTIC PROTEASE 37
2.2.1 PROTEASE DEPENDENT GROWTH ON PROTEINS AS SOLE NITROGEN SOURCE 37
2.2.1.1 GROWTH OF DIFFERENT SAP MUTANTS WITH BSA AS SOLE NITROGEN SOURCE
37
2.2.1.2 DSY459 IS HOMOZYGOUS FOR CHROMOSOME R 39
2.2.1.3 REINTRODUCTION OF THE SAP2-2 ALLELE RESTORED GROWTH OF DSY459 42
2.2.1.4 HOMOZYGOSITY FOR CHROMOSOME R AND SAP2-1 IS SUFFICIENT TO RESULT
IN A
GROWTH DEFECT WITH PROTEINS AS SOLE NITROGEN SOURCE 45
HTTP://D-NB.INFO/1044067608
2.2.1.5 LACK OF THE SAP2-2 ALLELE IS SUFFICIENT TO RESULT IN A GROWTH
DEFECT WITH
PROTEINS AS SOLE NITROGEN SOURCE 46
2.2.2 SAP2 PROMOTER DELETION ANALYSIS 49
2.2.2.1 DEFINING SAP2 REGULATORY SEQUENCES 50
2.2.2.2 SAP2 EXPRESSION ANALYSIS 51
2.2.2.3 SAP2-2 IS INDUCED EARLIER AND BETTER THAN SAP2-1 51
2.2.2.4 CONSTRUCTION OF DELETED SAP2-1 PROMOTERS 53
2.2.2.5 CONSTRUCTION DELETED SAP2-2 PROMOTERS 56
2.2.2.6 SAP2 EXPRESSION IS CONTROLLED BY DISTINCT PROMOTER ELEMENTS 59
2.2.2.7 SAP2 EXPRESSION IS INDUCED INDEPENDENT OF STPL 63
2.3 TRANSPORTER-MEDIATED PEPTIDE UPTAKE 65
2.3.1 GROWTH ON AMINO ACIDS AS SOLE NITROGEN SOURCE 66
2.3.2 CONTRIBUTION OF DIFFERENT TRANSPORTERS TO THE GROWTH WITH BSA AS
SOLE NITROGEN
SOURCE 67
2.3.3 CONTRIBUTION OF DIFFERENT TRANSPORTERS TO THE GROWTH WITH DEFINED
PEPTIDES AS
SOLE NITROGEN SOURCE 68
2.3.4 GROWTH RATE DETERMINATION 72
2.3.5 IMPACT OF PEPTIDE-UPTAKE DEFICIENCY ON IN VITRO FITNESS 73
2.3.6 IMPACT OF PEPTIDE-UPTAKE DEFICIENCY ON IN VIVO FITNESS 75
2.4 CONTROL OF THE EXPRESSION OF VIRULENCE-ASSOCIATED TRAITS BY AMMONIUM
77
2.4.1 VARIOUS AMMONIUM CONCENTRATIONS REPRESS SAP2 EXPRESSION TO VARIOUS
DEGREES
77
2.4.2 AMMONIUM UPTAKE IS REQUIRED TO REPRESS SAP2 EXPRESSION 79
2.4.3 AMMONIUM UPTAKE IS REQUIRED TO REPRESS OPT3 EXPRESSION 82
2.4.4 AMMONIUM UPTAKE IS REQUIRED TO REPRESS DALI AND DUR3 EXPRESSION 84
2.4.5 AMMONIUM UPTAKE IS REQUIRED TO REPRESS ARGININE-INDUCED
FILAMENTATION 86
3 DISCUSSION 88
3.1 STPL, A TRANSCRIPTIONAL REGULATOR INVOLVED IN PROTEIN UTILIZATION
AND BEYOND 88
3.2 ONLY SAP2 IS REQUIRED FOR GROWTH WITH BSA AS SOLE NITROGEN SOURCE
AND ITS EXPRESSION
IS CONTROLLED BY DISTINCT C/S-REGULATORY SEQUENCES 92
3.3 UTILIZATION OF PROTEINS AND PEPTIDES ARE NOT REQUIRED FOR C.
ALBICANS GUT COLONIZATION
99
3.4 AMMONIUM MUST BE TAKEN UP TO CONTROL THE EXPRESSION OF
VIRULENCE-ASSOCIATED TRAITS
103
4 MATERIAL AND METHODS 109
4.1 MATERIAL 109
4.1.1 GROWTH MEDIA AND AGAR 109
4.1.2 GENERAL SOLUTIONS 109
4.1.3 ENZYMES AND CHEMICALS 110
4.1.4 OLIGONUCLEOTIDES 112
4.1.5 PLASMIDS AND C. ALBICANS MUTANTS 114
4.1.5.1 PLASMIDS 115
4.1.5.2 C. ALBICANS STRAINS 118
4.2 METHODS 124
4.2.1 GROWTH OF ESCHERISCHA COLI STRAINS 124
4.2.2 GROWTH OF CANDIDA ALBICANS STRAINS 124
4.2.3 DNA MANIPULATION AND STRAIN CONSTRUCTION 124
4.2.3.1 AGAROSE GEL ELECTROPHORESIS AND VISUALIZATION OF DNA-FRAGMENTS
124
4.2.3.2 PCR (POLYMERASE CHAIN REACTION) 125
4.2.3.3 DIGESTION AND PURIFICATION OF DNA FRAGMENTS 125
4.2.3.4 PLASMID CONSTRUCTION 125
4.2.3.5 PREPARATION OF COMPETENT E. COLI CELLS 126
4.2.3.6 TRANSFORMATION OF COMPETENT E. COLI CELLS 126
4.2.3.7 ISOLATION OF E. COLI PLASMID DNA 126
4.2.3.8 SEQUENCING 127
4.2.3.9 TRANSFORMATION OF CANDIDA ALBICANS 127
4.2.3.10 ISOLATION OF GENOMIC DNA FROM C. ALBICANS 128
4.2.3.11 SOUTHERN DNA TRANSFER AND HYBRIDIZATION 128
4.2.4 EXPRESSION REPORTER ASSAYS 129
4.2.4.1 QUANTITATIVE REAL TIME PCR 129
4.2.4.2 FLOW CYTOMETRY 132
4.2.4.3 RAW PROTEIN EXTRACT ISOLATION 132
4.2.4.4 SDS-PAGE 132
4.2.4.5 WESTERN BLOTTING AND ANTIBODY DETECTION 133
4.2.4.6 ENZYME-IINKED IMMNUNOSORBENT ASSAY (ELISA) 133
4.2.5 PHENOTYPIC ASSAYS 134
4.2.5.1 GROWTH RATE DETERMINATION 134
4.2.5.2 IN VITRO COMPETITIVE FITNESS ASSAY 134
4.2.5.3 IN VIVO COMPETITIVE FITNESS ASSAY 135
4.2.6 GENOME-WIDE BINDING PROFILING (CHLP-ON-CHIP) 136
REFERENCES 138
APPENDIX : 148
APPENDIX AI: STPL TARGETS 148
APPENDIX A2:
SAP2 SEQUENCES FROM PLASMIDS PSAP2KS4 AND PSAP2KS5 155
|
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| institution | BVB |
| language | German |
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| spelling | Dunkel, Nico 1983- Verfasser (DE-588)104340466X aut Regulation of virulence-associated traits of the human fungal pathogen Candida albicans by nitrogen availability von Nico Dunkel Regulation Virulenz-assoziierter Faktoren im humanpathogenen Pilz Candida albicans durch Stickstoffverfügbarkeit 2013 IV, 158, IV S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. und engl. Sprache Würzburg, Univ., Diss., 2013 Candida albicans (DE-588)4147244-5 gnd rswk-swf Proteasen (DE-588)4047521-9 gnd rswk-swf Stickstoff (DE-588)4125639-6 gnd rswk-swf Regulation (DE-588)4049075-0 gnd rswk-swf Virulenz (DE-588)4129834-2 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Candida albicans (DE-588)4147244-5 s Virulenz (DE-588)4129834-2 s Regulation (DE-588)4049075-0 s Stickstoff (DE-588)4125639-6 s Proteasen (DE-588)4047521-9 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:20-opus-83076 https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-83076 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=026829547&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Dunkel, Nico 1983- Regulation of virulence-associated traits of the human fungal pathogen Candida albicans by nitrogen availability Candida albicans (DE-588)4147244-5 gnd Proteasen (DE-588)4047521-9 gnd Stickstoff (DE-588)4125639-6 gnd Regulation (DE-588)4049075-0 gnd Virulenz (DE-588)4129834-2 gnd |
| subject_GND | (DE-588)4147244-5 (DE-588)4047521-9 (DE-588)4125639-6 (DE-588)4049075-0 (DE-588)4129834-2 (DE-588)4113937-9 |
| title | Regulation of virulence-associated traits of the human fungal pathogen Candida albicans by nitrogen availability |
| title_alt | Regulation Virulenz-assoziierter Faktoren im humanpathogenen Pilz Candida albicans durch Stickstoffverfügbarkeit |
| title_auth | Regulation of virulence-associated traits of the human fungal pathogen Candida albicans by nitrogen availability |
| title_exact_search | Regulation of virulence-associated traits of the human fungal pathogen Candida albicans by nitrogen availability |
| title_full | Regulation of virulence-associated traits of the human fungal pathogen Candida albicans by nitrogen availability von Nico Dunkel |
| title_fullStr | Regulation of virulence-associated traits of the human fungal pathogen Candida albicans by nitrogen availability von Nico Dunkel |
| title_full_unstemmed | Regulation of virulence-associated traits of the human fungal pathogen Candida albicans by nitrogen availability von Nico Dunkel |
| title_short | Regulation of virulence-associated traits of the human fungal pathogen Candida albicans by nitrogen availability |
| title_sort | regulation of virulence associated traits of the human fungal pathogen candida albicans by nitrogen availability |
| topic | Candida albicans (DE-588)4147244-5 gnd Proteasen (DE-588)4047521-9 gnd Stickstoff (DE-588)4125639-6 gnd Regulation (DE-588)4049075-0 gnd Virulenz (DE-588)4129834-2 gnd |
| topic_facet | Candida albicans Proteasen Stickstoff Regulation Virulenz Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-83076 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026829547&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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