Analysis of the transcriptional regulation of the MOX gene encoding peroxisomal methanol oxidase from Hansenula polymorpha:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
1994
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | 100 Bl. graph. Darst. |
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| 100 | 1 | |a Pereira, Gonçalo Amarante Guimaraes |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Analysis of the transcriptional regulation of the MOX gene encoding peroxisomal methanol oxidase from Hansenula polymorpha |c vorgelegt von Gonçalo Amarante Guimaraes Pereira |
| 264 | 1 | |c 1994 | |
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| 337 | |b n |2 rdamedia | ||
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| 502 | |a Düsseldorf, Univ., Diss. | ||
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Datensatz im Suchindex
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| adam_text |
CONTENT
CONTENT
1. INTRODUCTION 1
1.1 TRANSCRIPTIONAL REGULATION OF PROTEIN-ENCODING GENES IN YEAST 1
1.2 PEROXISOMES AND GENES ENCODING PEROXISOMAL PROTEINS 9
2. MATERIAL AND METHODS 15
2.1 CHEMICALS 15
2.2 YEASTS 15
2.2.1 STRAINS 15
2.2.2 GROWTH CONDITIONS 15
2.3
E. COLI 16
2.3.1 STRAINS 16
2.3.2 GROWTH CONDITIONS 16
2.4 RECOMBINANT DNA TECHNIQUES 16
2.5 BASIC PLASMIDS 16
2.5.1 FOR CLONING AND AMPLIFICATION OF DNA FRAGMENTS 16
2.5.2 FOR EXPRESSION OF GST-ADR1 FUSION PROTEIN 16
2.5.3 FOR PROMOTER ANALYSIS IN
H. POFYMORPHA 16
2.5.4 FOR PROMOTER ANALYSIS IN
S. CEREVISIAE 16
2.5.5 FOR OVEREXPRESSION OF ADR1 IN
S. CEREVISIAE 16
2.5.6 FOR ISOLATION OF GENE SEQUENCES 17
2.6 PLASMID CONSTRUCTION 17
2.6.1 FOR ANALYSIS OF
MOX PROMOTER IN H. POFYMORPHA 17
2.6.2 FOR ANALYSIS OF
MOX PROMOTER IN S. CEREVISIAE 17
2.6.3 FOR ANALYSIS OF OTHER
H. POFYMORPHA PROMOTERS IN S. CEREVISIAE 17
2.6.4 FOR EXPRESSION OF FUSIONS GST-ADR1 IN
E. COLI 18
2.6.5 FOR ISOLATION OF PROMOTER FRAGMENTS TO PERFORM GEL SHIFT
EXPERIMENTS 18
2.6.6 FOR DISRUPTION OF THE GENE ADR1 18
2.6.7 FOR DISRUPTION OF THE GENE
MIG1 18
2.6.8 FOR OVEREXPRESSION OF
MIG1 19
2.7
E.COLI TRANSFORMATION 19
2.8 YEAST TRANSFORMATION 19
2.9 PREPARATION OF YEAST DNA 19
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/943350875
CONTENT
2.10 PREPARATION AND ELECTROPHORESIS OF YEAST RNA 19
2.11 PREPARATION OF
H. POLYMORPHA CHROMATIN 20
2.12 LABELLING OF DNA PROBES AND CONDITIONS FOR HYBRIDIZATION 20
2.13 DETERMINATION OF PROTEIN CONCENTRATION 20
2.14 PROTEIN CRUDE EXTRACT AND ENZYMATIC ACTIVITY 21
2.14.1 B-GALACTOSIDASE 21
2.14.2 METHANOL OXIDASE 21
2.14.3 CATALASE 21
2.14.4 0-LACTAMASE 21
2.15 EXPERIMENTS FOR DETERMINATION OF PROTEIN/DNA INTERACTION 22
2.15.1 PROTEIN EXTRACTS FOR DNA BINDING ESSAYS 22
2.15.2 DNA FRAGMENTS FOR BINDING ASSAYS 22
2.15.3 GEL SHIFT ANALYSIS - 22
2.15.4 DNASEL FOOTPRINTING 23
3. RESULTS 24
3.1 INVESTIGATION OF THE DEREPRESSION PROCESS OF THE
MOX PROMOTER 24
3.1.1 STRATEGY FOR ISOLATION OF MUTANTS IMPAIRED IN THE
MOX DEREPRESSION 24
3.1.2 CHARACTERIZATION OF Q1N MUTANTS DEFECTIVE IN THE MOX DEREPRESSION
26
3.1.3 TRANSCRIPTIONAL REGULATION OF MOX IN MQ1N STRAINS 28
3.1.4 ANALYSIS OF DELETIONS IN THE
MOX PROMOTER IN THE GENE FUSION MOX-FILAC 31
3.1.5 KINETICS OF THE SYNTHESIS OF PREMOX IN THE MUTANT MQ1N8 35
3.2 REGULATION OF THE GENES ENCODING PEROXISOMAL PROTEINS INVOLVED IN
THE METHANOL METABOLISM 36
3.2.1 TRANSCRIPTIONAL REGULATION OF
MOX, DAS AND CTA 36
3.2.2 IMPORTANCE OF THE COORDINATION BETWEEN
MOX AND CTA FOR
THE METHANOL METABOLISM 38
3.2.3 DEREPRESSION OF THE GENES
MOX, CTA AND DAS IN THE MUTANT MQ1N8 39
3.3 GEL SHIFT ANALYSIS WITH PROTEIN CRUDE EXTRACT FROM
H. POLYMORPHA 41
3.3.1 THE
MOX-B REGION 41
3.4 CHROMATIN STRUCTURE OF THE
MOX PROMOTER 42
3.4.1 POSITIONING OF NUCLEOSOMES IN THE STRAIN Q1N 42
3.4.2 POSITIONING OF THE NUCLEOSOMES IN WILD TYPE CELLS 47
3.5 HETEROLOGOUS
MOX EXPRESSION IN SACCHAROMYCES CEREVISIAE 48
3.5.1 REGULATED EXPRESSION OF THE
MOX PROMOTER IN S. CEREVISIAE 48
CONTENT
3.5.2 DELETION ANALYSIS OF THE
MOX PROMOTER IN S. CEREVISIAE 49
3.5.3 EFFECT OF THE REGULATORS ADR1 AND MIG1 ON THE REGULATION OF THE
MOX PROMOTER 50
3.5.4 IDENTIFICATION OF THE ADR1 RESPONSE REGION IN THE
MOX
PROMOTER 51
3.5.5 ANALYSIS OF THE
MOX PROMOTER REGIONS IN HYBRID MOX-CYC1
PROMOTERS 53
3.5.6 EXPRESSION IN
S. CEREVISIAE OF OTHER H. POFYMORPHA
GENES INVOLVED IN METHANOL METABOLISM 54
3.5.7 BINDING OF ADR1 TO THE B REGION FROM THE
MOX PROMOTER 55
3.5.8 TWO MOLECULES OF ADR1 BIND TO THE
MOX-B REGION 56
3.5.9 COMPARISON BETWEEN THE BINDING OF FB2 AND ADR1 TO THE
MOX-B2 REGION 58
3.5.10 THE BINDING OF FB2 BENDS THE DNA 60
4. DISCUSSION 62
4.1 THE PROCESS OF
MOX DEREPRESSION IN H. POFYMORPHA 62
4.1.1 ISOLATION OF REGULATORY MUTANTS 62
4.1.2 THE PREMOX TRANSCRIPT AND ITS RELATION WITH THE
MOX DEREPRESSION 63
4.1.3 DIE DEREPRESSION OF MOX DEPENDS ON A TRANSCRIPTION START
POINT SWITCH MECHANISM 65
4.1.4 THE IMPORTANCE OF THE COORDINATION BETWEEN THE EXPRESSION
OF GENES ENCODING PEROXISOMAL PROTEINS 67
4.1.5 PREMOX AND THE COORDINATION OF THE DEREPRESSION OF THE GENES
MOX AND CTA 69
4.1.6 THE
MOX PROMOTER CHROMATIN AND ITS CONNECTION WITH
PREMOX AND THE TSP SWITCH 71
4.2 EXPRESSION OF
MOX IN S. CEREVISIAE 76
4.2.1 THE REGULATION OF
MOX IN S. CEREVISIAE IS UNIQUE AMONG
THE MAJOR GENES INVOLVED IN THE METHANOL METABOLISM 76
4.2.2 THE
MOX REGULATION IN S. CEREVISIAE AND THE ROLE OF ADR1 78
4.2.3 PHYSICAL INTERACTION BETWEEN ADR 1 AND THE MOX PROMOTER 80
4.3 PERSPECTIVES 83
5. SUMMARY/ZUSAMMENFASSUNG 84
6. REFERENCES 88
7. ABBREVIATIONS 100 |
| any_adam_object | 1 |
| author | Pereira, Gonçalo Amarante Guimaraes |
| author_facet | Pereira, Gonçalo Amarante Guimaraes |
| author_role | aut |
| author_sort | Pereira, Gonçalo Amarante Guimaraes |
| author_variant | g a g p gag gagp |
| building | Verbundindex |
| bvnumber | BV010081779 |
| classification_tum | CHE 832d BIO 325d BIO 180d BIO 220d |
| ctrlnum | (OCoLC)64521226 (DE-599)BVBBV010081779 |
| discipline | Biologie Chemie |
| format | Thesis Book |
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| indexdate | 2024-10-09T16:18:35Z |
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| language | English |
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| physical | 100 Bl. graph. Darst. |
| publishDate | 1994 |
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| spelling | Pereira, Gonçalo Amarante Guimaraes Verfasser aut Analysis of the transcriptional regulation of the MOX gene encoding peroxisomal methanol oxidase from Hansenula polymorpha vorgelegt von Gonçalo Amarante Guimaraes Pereira 1994 100 Bl. graph. Darst. txt rdacontent n rdamedia nc rdacarrier Düsseldorf, Univ., Diss. Genregulation (DE-588)4122166-7 gnd rswk-swf Gen (DE-588)4128987-0 gnd rswk-swf Methanoloxidase (DE-588)4363991-4 gnd rswk-swf Hansenula polymorpha (DE-588)4159091-0 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Hansenula polymorpha (DE-588)4159091-0 s Methanoloxidase (DE-588)4363991-4 s Genregulation (DE-588)4122166-7 s DE-604 Gen (DE-588)4128987-0 s DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=006691342&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Pereira, Gonçalo Amarante Guimaraes Analysis of the transcriptional regulation of the MOX gene encoding peroxisomal methanol oxidase from Hansenula polymorpha Genregulation (DE-588)4122166-7 gnd Gen (DE-588)4128987-0 gnd Methanoloxidase (DE-588)4363991-4 gnd Hansenula polymorpha (DE-588)4159091-0 gnd |
| subject_GND | (DE-588)4122166-7 (DE-588)4128987-0 (DE-588)4363991-4 (DE-588)4159091-0 (DE-588)4113937-9 |
| title | Analysis of the transcriptional regulation of the MOX gene encoding peroxisomal methanol oxidase from Hansenula polymorpha |
| title_auth | Analysis of the transcriptional regulation of the MOX gene encoding peroxisomal methanol oxidase from Hansenula polymorpha |
| title_exact_search | Analysis of the transcriptional regulation of the MOX gene encoding peroxisomal methanol oxidase from Hansenula polymorpha |
| title_full | Analysis of the transcriptional regulation of the MOX gene encoding peroxisomal methanol oxidase from Hansenula polymorpha vorgelegt von Gonçalo Amarante Guimaraes Pereira |
| title_fullStr | Analysis of the transcriptional regulation of the MOX gene encoding peroxisomal methanol oxidase from Hansenula polymorpha vorgelegt von Gonçalo Amarante Guimaraes Pereira |
| title_full_unstemmed | Analysis of the transcriptional regulation of the MOX gene encoding peroxisomal methanol oxidase from Hansenula polymorpha vorgelegt von Gonçalo Amarante Guimaraes Pereira |
| title_short | Analysis of the transcriptional regulation of the MOX gene encoding peroxisomal methanol oxidase from Hansenula polymorpha |
| title_sort | analysis of the transcriptional regulation of the mox gene encoding peroxisomal methanol oxidase from hansenula polymorpha |
| topic | Genregulation (DE-588)4122166-7 gnd Gen (DE-588)4128987-0 gnd Methanoloxidase (DE-588)4363991-4 gnd Hansenula polymorpha (DE-588)4159091-0 gnd |
| topic_facet | Genregulation Gen Methanoloxidase Hansenula polymorpha Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=006691342&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT pereiragoncaloamaranteguimaraes analysisofthetranscriptionalregulationofthemoxgeneencodingperoxisomalmethanoloxidasefromhansenulapolymorpha |