Molecular characterisation of three novel photosynthetic proteins in arabidopsis thaliana:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2008
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| Schlagworte: | |
| Online-Zugang: | kostenfrei https://nbn-resolving.org/urn:nbn:de:bvb:19-163720 Inhaltsverzeichnis |
| Beschreibung: | VIII, 109 Bl. graph. Darst. |
Internformat
MARC
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| 035 | |a (OCoLC)867157121 | ||
| 035 | |a (DE-599)BVBBV041462290 | ||
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| 100 | 1 | |a Armbruster, Ute |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Molecular characterisation of three novel photosynthetic proteins in arabidopsis thaliana |c von Ute Armbruster |
| 264 | 1 | |c 2008 | |
| 300 | |a VIII, 109 Bl. |b graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a München, Univ., Diss., 2008 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
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| 856 | 4 | 1 | |u http://edoc.ub.uni-muenchen.de/16372/ |x Verlag |z kostenfrei |3 Volltext |
| 856 | 4 | |u https://nbn-resolving.org/urn:nbn:de:bvb:19-163720 |x Resolving-System | |
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| 912 | |a ebook | ||
| 999 | |a oai:aleph.bib-bvb.de:BVB01-026908617 | ||
Datensatz im Suchindex
| _version_ | 1804151592979529728 |
|---|---|
| adam_text | CONTENTS
CONTENTS
SUMMARY I
ZUSAMMENFASSUNG II
CONTENTS III
ABBREVIATIONS VII
1 INTRODUCTION 1
1.1 PHOTOSYNTHESIS 1
1.1.1 PHOTOSYNTHETIC ELECTRON TRANSPORT 1
1.1.1.1 LINEAR ELECTRON TRANSPORT 1
1.1.1.2 CYCLIC ELECTRON TRANSPORT AROUND PSI 2
1.1.1.3 PHYSICAL MEASUREMENTS OF ELECTRON TRANSPORT 4
1.1.2 CARBON FIXATION 4
1.1.3 THE THYLAKOID MEMBRANE 5
1.1.3.1 THE THYLAKOID POLYPEPTIDE COMPOSITION 6
1.1.4 REGULATION OF PHOTOSYNTHETIC PROCESSES 8
1.1.4.1 ENERGISATION DEPENDENT NON-PHOTOCHEMICAL QUENCHING (QE) 8
1.1.4.2 STATE TRANSITIONS 9
1.2 EXPRESSION OF PHOTOSYNTHETIC GENES 9
1.2.1 NUCLEAR GENE EXPRESSION 10
1.2.1 CHLOROPLAST GENE EXPRESSION 11
1.3 ABSCISIC ACID (ABA) BIOSYNTHESIS IN PLANTS 11
1.4 AIMS OF THESIS 12
2 MATERIALS AND METHODS 14
2.1 MATERIALS 14
2.1.1 CHEMICALS 14
2.1.2 ANTIBIOTICS 14
2.1.3 ENZYMES, KITS AND BIOCHEMICAL AGENTS 14
2.1.4 MEMBRANES : 14
2.1.5 ANTIBODIES 14
2.2 METHODS IS
2.2.1 PLANT LINES AND PROPAGATION 15
III
HTTP://D-NB.INFO/1046876619
CONTENTS
2.2.2 NUCLEIC ACID ANALYIS 15
2.2.2.1 DNA ANALYSIS 15
2.2.2.2 RNA ANALYSIS 17
2.2.2.3 ANALYSIS OF MRNAS ASSOCIATED WITH POLYSOMES 17
2.2.2.4 TRANSCRIPT END MAPPING OF RBCL USING CIRCULAR RT-PCR 18
2.2.3 TRANSFORMATION OF ARABIDOPSIS 18
2.2.3.1. BACTERIAL STRAINS 18
2.2.3.2 AGROBACTERIUM BINARY VECTORS 18
2.2.3.3 AGROBACTERIUM-MEDIATED TRANSFORMATION OF A. THALICMA 19
2.2.4 BIOCHEMICAL ANALYSIS 19
2.2.4.1 ANTIBODY PRODUCTION 19
2.2.4.2 SDS-PAGE 19
2.2.4.3 IMMUNOBLOT ANALYSIS 19
2.2.4.4 TOTAL PROTEIN ISOLATION 20
2.2.4.5 ISOLATION OF INTACT CHLOROPLASTS 20
2.2.4.6 FRACTIONATION OF CHLOROPLASTS 20
2.2.4.7 PREPARATION OF THYLAKOID MEMBRANES 20
2.2.4.8 FRACTIONATION OF THYLAKOIDS 21
2.2.4.9 PSI ISOLATION 21
2.2.4.10 BLUE NATIVE AND SECOND DIMENSION GELS 21
2.2.4.11 BIS (SULFOSUCCINIMIDYL) SUBERATE (BS
3
) CROSSLINKING 22
2.2.4.12 CO-IMMUNOPRECIPITATION 22
2.2.4.13 SALT TREATMENT OF THYLAKOID MEMBRANES 22
2.2.4.14 TRYPSIN TREATMENT OF THYLAKOID MEMBRANES 23
2.2.4.15 IN VIVO LABELLING WITH
35
S-METHIONINE 23
2.2.5 MASS SPECTROMETRY : 23
2.2.5.1 TRYPTIC IN GEL DIGESTION OF PROTEINS 23
2.2.5.2 LC-ESI MS/MS 24
2.2.5.3 PROTEIN IDENTIFICATION 24
2.2.6 PIGMENT ANALYSIS 24
2.2.7 DATABASE ANALYSIS, DIGITAL NORTHERN, PREDICTION OF SUBCELLULAR
TARGETING AND PROTEIN
MODELLING 25
2.2.8 INTRACELLULAR LOCALIZATION OF DSRED FUSIONS 25
IV
CONTENTS
2.2.9 GERMINATION ASSAY 26
2.2.10 DETERMINATION OF PHOTOSYNTHETIC PARAMETERS USING THE PAM
FLUOROMETER 26
2.2.10.1 CHLOROPHYLL FLUORESCENCE MEASUREMENTS 26
2.2.10.2 MEASUREMENTS OF THE REDOX STATE OF P700 26
2.2.10.3 CYCLIC ELECTRON FLOW MEASUREMENTS 27
3 RESULTS 28
3.1 CHARACTERISATION OF PPP4 AND TMP14 28
3.1.1 DESCRIPTION OF A NOVEL PROTEIN FAMILY IN PHOTOSYNTHETIC ORGANISMS
28
3.1.2 LOCALISATION OF PPP4 AND TMP14 IN THE THYLAKOID MEMBRANE 31
3.1.3 KNOCK-OUT MUTANTS OF PPP4 AND TMP14 32
3.1.4 LOCALISATION OF PPP4 IN BN-PAGE 34
3.1.5 TMP14 PROTEIN LEVELS ARE DECREASED INPPP4 MUTANTS 37
3.1.6 INVESTIGATING THE INTERACTION BETWEEN PPP4 AND TMP14 37
3.1.6.1 EPITOPE TAGGING OFPPP4 37
3.1.6.2 CROSSLINKING 38
3.1.6.3 CO-IMMUNOPRECIPITATION (CO-IP) 41
3.1.7 TMP14 AND PPP4 ARE NOT STABLY ASSOCIATED WITH ANY OF THE MAIN
PHOTOSYNTHETIC COMPLEXES43
3.1.8 PHOTOSYNTHETIC PARAMETER OF WT, PPP4, TMPL4,PPP4 TMPL4 AND
PPP4:C-MYC
OVEREXPRESSORS 45
3.1.9 TOPOLOGY OF PPP4 AND TMP14 49
3.1.10 ANALYSIS OF THYLAKOID PROTEIN COMPOSITION 52
3.1.11 LOCATION OF THYLAKOID PROTEINS IN WT AND PPP4 TMPL4 53
3.1.12 INVESTIGATING PHOSPHORYLATION OF THYLAKOID PROTEINS IN WT AND
PPP4 TMPL4 55
3.1.13 CHARACTERISATION OF PPP4-LIKE AND TMP14-LIKE 55
3.2 CHARACTERISATION OF PPP1 58
3.2.1 GENE EXPRESSION, PROTEIN STRUCTURE AND LOCALISATION 58
3.2.2 PHENOTYPICAL ANALYSIS OF KNOCK-OUT MUTANTS OF PPP1 AND CSP41 60
3.2.3 LEAF PIGMENTS AND PHOTOSYNTHETIC PARAMETERS 62
3.2.4 ANALYSIS OF THE ABUNDANCE OF PHOTOSYNTHETIC PROTEINS :....
65
3.2.5 CHLOROPLAST EXPRESSION ANALYSIS 67
3.2.6 ANALYSIS OF TRANSCRIPT PROCESSING 68
3.2.7 EFFECT OF THE ABSENCE OF PHOTOSYNTHETIC COMPLEXES ON PPP1
ABUNDANCE 70
V
CONTENTS
3.2.8 A PPP1
:CFP FUSION COMPLEMENTS THEPPPL MUTANT PHENOTYPE 70
3.2.9 GERMINATION ASSAY 71
4 DISCUSSION 73
4.1 PPP4/TMP14 73
4.1.1 NOVEL FAMILY OF CHLOROPLAST LOCALISED PROTEINS 73
4.1.2 A NOVEL THYLAKOID COMPLEX PREDOMINANTLY LOCALISED IN THE STROMAL
LAMELLAE 73
4.1.3 PHENOTYPE OF PPP4 AND PPP4 TMPL4 MUTANTS IN RELATION TO SPECIFIC
PROCESSES OF
PHOTOSYNTHESIS 75
4.1.3.1 LINEAR ELECTRON FLOW 75
4.1.3.2 REGULATORY PROCESSES IN PHOTOSYNTHESIS 76
4.1.3.3 CYCLIC ELECTRON FLOW AROUND PSI (CEF) 77
4.1.3.3.1 IN-DEPTH ANALYSIS OF CEF IN PPP4 ANDPPP4 TMPL4 MUTANTS 79
4.1.3.3.2 SPECULATION ABOUT AN INVOLVEMENT OF PPP4 AND TMP14 IN CEF
PROCESSES 80
4.1.4 CONCLUSIONS 81
4.2 THE PUTATIVE RNA-BINDING PROTEIN PPP1 82
4.2.1 PHYSIOLOGICAL ROLE: CHLOROPLAST GENE EXPRESSION 82
4.2.2 THE HOMOLOGUE ATCSP41 84
4.2.3 INVOLVEMENT OF PPP1 IN ABA SIGNALLING 84
5 REFERENCES 87
6 SUPPLEMENTARY DATA 101
6.1 MASS SPECTROMETRY FRAGMENTATION SPECTRA 101
6.1.1 PPP4 101
6.1.2 TMP14 103
6.1.3 TMP14-LIKE 104
6.2 PLASTOME EXPRESSION ANALYSIS OF
PPPL
MUTANTS 105
ACKNOWLEDGEMENTS 106
CURRICULUM VITAE 107
EHRENWORTLICHE VERSICHERUNG 109
VI
|
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| spelling | Armbruster, Ute Verfasser aut Molecular characterisation of three novel photosynthetic proteins in arabidopsis thaliana von Ute Armbruster 2008 VIII, 109 Bl. graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Univ., Diss., 2008 (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:19-163720 http://edoc.ub.uni-muenchen.de/16372/ Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:19-163720 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026908617&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Armbruster, Ute Molecular characterisation of three novel photosynthetic proteins in arabidopsis thaliana |
| subject_GND | (DE-588)4113937-9 |
| title | Molecular characterisation of three novel photosynthetic proteins in arabidopsis thaliana |
| title_auth | Molecular characterisation of three novel photosynthetic proteins in arabidopsis thaliana |
| title_exact_search | Molecular characterisation of three novel photosynthetic proteins in arabidopsis thaliana |
| title_full | Molecular characterisation of three novel photosynthetic proteins in arabidopsis thaliana von Ute Armbruster |
| title_fullStr | Molecular characterisation of three novel photosynthetic proteins in arabidopsis thaliana von Ute Armbruster |
| title_full_unstemmed | Molecular characterisation of three novel photosynthetic proteins in arabidopsis thaliana von Ute Armbruster |
| title_short | Molecular characterisation of three novel photosynthetic proteins in arabidopsis thaliana |
| title_sort | molecular characterisation of three novel photosynthetic proteins in arabidopsis thaliana |
| topic_facet | Hochschulschrift |
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