Regulation of BBX24 and BBX25 proteins in Arabidopsis thaliana:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | VI, 133 Bl. Ill., graph. Darst. 30 cm 1 CD-R |
Internformat
MARC
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Datensatz im Suchindex
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| adam_text | TABLE OF CONTENTS
ABBREVIATIONS V
1. INTRODUCTION 1
1.1 PHOTORECEPTORS AND LIGHT-MEDIATED DEVELOPMENT 1
1.2 THE 26S PROTEASOME DEGRADATION PATHWAY. 5
1.3 COP/DET/FUS PROTEINS REGULATE PHOTOMORPHOGENESIS .7
1.4 HY5, A POSITIVE KEY REGULATOR OF PHOTOMORPHOGENESIS 10
1.5 THE ARABIDOPSIS THALIANA B-BOX (BBX) ZINC FINGER PROTEIN FAMILY 11
1.5.1 BBX24 AND BBX25 13
1.6 UV-B RADIATION 15
1.6.1 THE UV-B INDUCED PHOTOMORPHOGENESIS. 16
1.7 AIM OF THE THESIS. 20
2. MATERIAL AND METHODS 21
2.1 MATERIALS. 21
2.1.1 CHEMICALS 21
2.1.2 EQUIPMENT AND INSTRUMENTATION 21
2.1.3 ENZYMES 22
2.1.4 CULTURE MEDIA 22
2.1.4.1 BACTERIAL MEDIA 22
2.1.4.2 ARABIDOPSIS MURASHIGE & SKOOG (MS) MEDIA 23
2.1.5 NUCLEIC ACIDS 23
2.1.5.1 OLIGONUCLEOTIDES 23
2.1.5.2 VECTORS 25
2.1.5.3 PROBES FOR NORTHERN BLOT ANALYSIS 25
2.1.6 ANTIBODIES. 26
2.1.7 BACTERIA STRAINS 27
2.1.8 PLANT MATERIAL. 27
2.1.9 SOFTWARE AND DATABASES 28
HTTP://D-NB.INFO/1044358815
2.2 METHODS. 29
2.2.1 DNA ANALYSIS AND MANIPULATION 29
2.2.1.1 DNA ENZYMATIC TREATMENTS. 29
2.2.1.2 CONSTRUCTION OF THE BBX25-C
:HIS6X
EXPRESSION VECTOR 29
2.2.1.3 SEPARATION OF DNA FRAGMENTS ON AGAROSE GELS 29
2.2.1.4 SMALL- AND MEDIUM-SCALE PLASMID DNA PREPARATION 30
2.2.1.5 ISOLATION OF GENOMIC DNA FROM PLANTS 30
2.2.1.6 POLYMERASE CHAIN REACTION (PCR) 31
2.2.2 RNA METHODS 31
2.2.2.1 RNA EXTRACTION AND PURIFICATION FOR QRT-PCR ANALYSIS 31
2.2.2.2 RNA EXTRACTION FOR NORTHERN BLOT ANALYSIS 32
2.2.2.3 RNA QUANTIFICATION 32
2.2.2.4 SEPARATION OF RNA FRAGMENTS ON MOPS-FORMALDEHYDE GELS 33
2.2.2.5 QRT-PCR ANALYSIS 33
2.2.2.6 NORTHERN BLOT ANALYSIS 34
2.2.3 PROTEIN METHODS .35
2.2.3.1 EXPRESSION OF GST AND HIS TAG FUSION PROTEINS IN E. COLI. 35
2.2.3.2 PREPARATION OF CRUDE BACTERIAL CELL EXTRACTS 36
2.2.3.3 AFFINITY PURIFICATION OF RECOMBINANT PROTEINS FROM E. COLI 36
2.2.3.3.1 NATIVE PURIFICATION OF GST:BBX24-C FUSION PROTEIN 36
2.2.3.3.2 PURIFICATION OF BBX25-C:HIS
6X
UNDER DENATURING CONDITIONS .37
2.2.3.4 AFFINITY PURIFICATION OF BBX24- AND BBX25-SPECIFIC ANTIBODIES 38
2.2.3.5 DOT BLOT ANALYSIS. 39
2.2.3.6 PROTEIN PREPARATIONS FROM PLANT TISSUES 40
2.2.3.7 PROTEIN QUANTIFICATION 41
2.2.3.8 SEPARATION OF PROTEINS BY SDS-PAGE 41
2.2.3.9 PROTEIN BAND STAINING IN POLYACRYLAMIDE GELS 43
2.2.3.10 IMMUNOBLOT ANALYSIS .43
2.2.3.11 CO-IMMUNOPRECIPITATION AND MASS SPECTROMETRY ANALYSIS 45
2.2.3.12 TANDEM AFFINITY PURIFICATION OF PLANT PROTEINS ...47
2.2.4 MICROBIOLOGICAL METHODS 48
2.2.4.1 TRANSFORMATION OF ELECTRO-COMPETENT E. COLI. 48
2.2.4.2 PREPARATION AND TRANSFORMATION OF CHEMO-COMPETENT E. COLI 49
2.2.4.3 TRANSFORMATION OF AGROBACTERIUM TUMEFACIENS. 49
2.2.5 PLANT METHODS 50
2.2.5.1 SURFACE STERILIZATION OF ARABIDOPSIS SEEDS 50
2.2.5.2 ARABIDOPSIS GROWTH CONDITIONS ON MS MEDIA 50
2.2.5.3 ARABIDOPSIS GROWTH CONDITIONS ON SOIL 51
2.2.5.4 SELECTION OF BASTA RESISTANT PLANTS ON SOIL 51
2.2.5.5 ARABIDOPSIS TRANSFORMATION VIA FLORAL-DIP METHOD 51
2.2.6 LIGHT TREATMENTS OF PLANTS 52
2.2.6.1 CWL AND MONOCHROMATIC LIGHT TREATMENTS 52
2.2.6.2 UV-B LIGHT TREATMENTS 53
2.2.7 LIGHT INTENSITY MEASUREMENTS. 54
2.2.8 PHENOTYPIC ANALYSIS 54
2.2.9 PLANT HYPOCOTYL MEASUREMENTS 55
2.2.10 QUANTIFICATION OF ANTHOCYANIN AMOUNTS 56
2.2.11 EPI-FLUORESCENCE MICROSCOPY ANALYSIS OF PLANTS 56
3. RESULTS .57
3.1 POLYCLONAL ANTIBODIES GENERATION 57
3.1.1 GENERATION OF POLYCLONAL ANTIBODIES SPECIFIC TO BBX24 AND BBX25 57
3.1.2 SPECIFICITY OF BBX24- AND BBX25-ANTISERA 59
3.2 REGULATION OF BBX24 AND BBX25 IN RESPONSE TO VISIBLE LIGHT 63
3.2.1 BBX24 AND BBX25 ACCUMULATE DURING SEEDLING DE-ETIOLATION 63
3.2.2 BLUE AND FAR-RED LIGHT PROMOTE THE ACCUMULATION OF BBX24 66
3.2.3 COP1 IS REQUIRED FOR BBX24 AND BBX25 TURNOVER IN LIGHT 68
3.2.4 HY5 IS REQUIRED FOR BBX24 AND BBX25 TURNOVER IN LIGHT 71
3.2.5 RELATIONSHIP BETWEEN BBX24 AND RCD1, AND SR01 73
3.2.6 DIURNAL REGULATION OF BBX24 EXPRESSION IS DEPENDENT ON HY5 74
3.2.7 DET1 REGULATES THE EXPRESSION OF BBX24 IN DARKNESS AND IN LIGHT 77
3.3 BBX24 AND BBX25 REGULATION IN THE RESPONSE TO UV-B LIGHT .78
3.3.1 FUNCTIONAL CHARACTERIZATION OF BBX24 IN THE RESPONSE TO UV-B LIGHT
78
3.3.2 UV-B PROMOTES ACCUMULATION OF BBX24 80
3.3.3 UV-B INDUCED ACCUMULATION OF BBX24 IS DEPENDENT ON UVR8 AND
COP1 81
3.3.4 BROADBAND UV-B PROMOTES ACCUMULATION OF BBX24 AND BBX25 86
III
3.3.5 UVR8 IS REQUIRED FOR THE BROADBAND UV-B INDUCED ACCUMULATION OF
BBX24, BUT NOT COP1 AND HY5. 89
3.4 BBX24, BBX25, AND RCD1 ARE INVOLVED IN LIGHT SIGNALLING IN LOW WL
INTENSITIES. .91
3.5 APPROACHES TO IDENTIFY BBX24 INTERACTION PARTNERS 93
4. DISCUSSION .98
5. SUMMARY. 109
6. REFERENCES. 112
7. SUPPLEMENTARY MATERIAL
(CD IN THE INSIDE BACK COVER)
7.1 DESCRIPTION OF THE VECTORS USED IN THIS STUDY
7.2 SUPPLEMENTAL FIGURES
|
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| author | Marquardt, Katrin |
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| discipline | Biologie Medizin |
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| spelling | Marquardt, Katrin Verfasser aut Regulation of BBX24 and BBX25 proteins in Arabidopsis thaliana presented by Katrin Marquardt 2013 VI, 133 Bl. Ill., graph. Darst. 30 cm 1 CD-R txt rdacontent n rdamedia nc rdacarrier Freiburg (Breisgau), Univ., Diss., 2013 (DE-588)4113937-9 Hochschulschrift gnd-content DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027010112&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Marquardt, Katrin Regulation of BBX24 and BBX25 proteins in Arabidopsis thaliana |
| subject_GND | (DE-588)4113937-9 |
| title | Regulation of BBX24 and BBX25 proteins in Arabidopsis thaliana |
| title_auth | Regulation of BBX24 and BBX25 proteins in Arabidopsis thaliana |
| title_exact_search | Regulation of BBX24 and BBX25 proteins in Arabidopsis thaliana |
| title_full | Regulation of BBX24 and BBX25 proteins in Arabidopsis thaliana presented by Katrin Marquardt |
| title_fullStr | Regulation of BBX24 and BBX25 proteins in Arabidopsis thaliana presented by Katrin Marquardt |
| title_full_unstemmed | Regulation of BBX24 and BBX25 proteins in Arabidopsis thaliana presented by Katrin Marquardt |
| title_short | Regulation of BBX24 and BBX25 proteins in Arabidopsis thaliana |
| title_sort | regulation of bbx24 and bbx25 proteins in arabidopsis thaliana |
| topic_facet | Hochschulschrift |
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