Isolation and molecular characterization of cDNAs encoding PRH75, a nucleolar DEAD-box protein from higher plants:
Gespeichert in:
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
1997
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | V, 106 S. Ill., graph. Darst. |
Internformat
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| 245 | 1 | 0 | |a Isolation and molecular characterization of cDNAs encoding PRH75, a nucleolar DEAD-box protein from higher plants |c von Zdravko Lorkovic |
| 264 | 1 | |c 1997 | |
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| 502 | |a München, Univ., Diss., 1997 | ||
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Datensatz im Suchindex
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| adam_text | ISOLATION AND MOLECULAR CHARACTERIZATION OF CDNAS ENCODING PRH75, A
NUDEOLAR DEAD-BOX PROTEIN FROM HIGHER PLANTS INAUGURAL-DISSERTATION ZUR
ERIANGUNG DES DOKTORGRADES DER FAKULTAT FUR BIOLOGIE DER
LUDWIG-MAXIMILIANS-UNIVERSITAT MIINCHEN VORGELEGT VON ZDRAVKO LORKOVIC
AUS JARCE POLJE, KROATIEN 1997 CONTENTS CONTENTS ABBREVIATIONS V SUMMARY
L I. INTRODUCTION 3 1.1. RNA STRUCTURE AND RNA RECOGNITION 4 1.2.
PROTEINS AND PROTEIN MOTIFS INVOLVED IN RNA RECOGNITION 5 1.3. A FAMILIY
OF RNA HELICASES 8 1.3.1. THE FUNCTION OF THE CONSERVED MOTIFS 10 1.3.2.
SPECIFICITY OF DEAD/H-BOX PROTEINS 11 N. MATERIALS AND METHODS 13 2.1.
MATERIALS 13 2.1. 1. CHEMICALS 13 2.1. 2. ENZYMES 13 2.1. 3. DNA LENGTH
STANDARDS 13 2.1. 4. PROTEIN MOLECULAR WEIGHT STANDARDS 13 2.1. 5.
CHROMATOGRAPHIC MEDIA 13 2.1. 6. ANTIBODIES 14 2.1. 7. BACTERIAL STRAINS
14 2.1. 8. PHAGES 14 2.1. 9. PLASMIDS 14 2.1.10. OLIGONUCLEOTIDES 15
2.1.11. PLANT MATERIAL 16 2.1.12. MEDIA FOR GROWTH OF BACTERIA AND
PHAGES 16 2.1.13. GENERAL BUFFERS AND SOLUTIONS 17 CONTENTS 2.2. METHODS
18 2.2.1. GENERAL MOLECULAR BIOLOGICAL METHODS 18 2.2.2. METHODS FOR DNA
MANIPULATION AND ANLYSIS 19 2.2.2.1. ISOLATION OF TOTAL PLANT DNA 19
2.2.2.2. SCREENING OF XGTL 1 CDNA LYBRARY (BENTON AND DAVIS, 1977) 20
2.2.2.3. ISOLATION OF PHAGE DNA 21 2.2.2.4. RADIOACTIVE LABELLING OF DNA
22 2.2.3. METHODS FOR RNA ISOLATION AND ANALYSIS 22 2.2.3.1. ISOLATION
OF PLANT TOTAL RNA AND NORTHERN BLOT ANALYSES 22 2.2.3.2. IN VITRO
TRANSCRIPTION 23 2.2.4. PROTEIN ANALYSIS 23 2.2.4.1. GENERAL METHODS FOR
PROTEIN ANALYSIS 23 2.2.4.2. OVEREXPRESSION OF RECOMBINANT PROTEINS IN
E. COLI 23 2.2.4.2.1. CONSTRUCTION OF PLASMIDS EXPRESSING THE PRH75
DELETION MUTANTS 23 2.2.4.2.2. CONSTRUCTION OF GST-PRH75 FUSIONS 24
2.2.4.2.3. OVEREXPRESSION OF PRH75 DELETION MUTANTS AND GST-PRH75 FUSION
PROTEINS 24 2.2.4.2.4. PURIFICATION OF GST-PRH75 FUSION PROTEINS 25
2.2.4.3. PROTEIN-RNA INTERACTION STUDIES (NORTHWESTERN ASSAY) 25
2.2.4.4. ATP HYDROLYSIS ASSAY 26 2.2.4.5. PREPARATION OF POLYCLONAL
ANTI-PRH75 ANTIBODIES 26 2.2.4.6. PREPARATION OF SPINACH WHOLE CELL
EXTRACT 27 2.2.4.6.1. FRACTIONATION OF SPINACH WHOLE CELL EXTRACTS ON
GLYCEROL GRADIENTS AND BY GEL FILTRATION 28 2.2.5. CELL BIOLOGY METHODS
28 2.2.5.1. CONSTRUCTION OF PLASMIDS FOR PROTOPLAST TRANSFORMATION 28
2.2.5.2. PROTOPLAST ISOLATION, TRANSFORMATION AND GUS ASSAY 29 2.2.5.3.
ISOLATION OF SPINACH*NUCLEI AND PREPARATION OF NUCLEAR PROTEIN EXTRACTS
30 2.2.5.4. FRACTIONATION OF NUCLEI INTO NUCLEOPLASM AND NUCLEOLI 31
CONTENTS 2.2.5.5. INDIRECT IMMUNOFLUORESCENCE ON ISOLATED SPINACH NUCLEI
32 2.2.5.6. CELLULAR FRACTIONATION 32 2.2.6. GENERATION OF TRANSGENIC
ARABIDOPSIS PLANTS 32 2.2.6.1. CONSTRUCTION OF PLASMIDS FOR ARABIDOPSIS
TRANSFORMATION 32 2.2.6.2. TRANSFORMATION OF A. THALIANA 33 III. RESULTS
34 3.1. ISOLATION OF GENES FOR REGULATORY NUCLEAR PROTEINS 34 3.2. CDNA
CLONING FOR AN DEAD-BOX PROTEIN FROM SPINACH 37 3.2.1. CDNA CLONING AND
PRIMARY STRUCTURE OF PRH75 37 3.2.2. PRH75 AND OTHER DEAD-BOX PROTEINS
42 3.2.3. EXPRESSION OF PRH75 DURING PLANT DEVELOPMENT 44 3.2.4. IN
VITRO RNA-BINDING STUDIES 46 3.2.4.1. PRH75 CONTAINS TWO RNA BINDING
SITES 46 3.2.4.2. THE C-TERMINAL GYR DOMAIN IS AN INDEPENDENT RNA-
BINDING MODULE 47 3.2.4.3. DETERMINATION OF THE SECOND RNA BINDING
DOMAIN IN PRH75 49 3.2.5. SUBCELLULAR LOCALIZATION OF PRH75 51 3.2.5.1.
CELLULAR FRACTIONATION AND TRANSIENT EXPRESSION OF PRH75::GUS FUSION IN
TOBACCO PROTOPLASTS DEMONSTRATE THE NUCLEAR LOCALIZATION OF PRH75 51
3.2.5.2. REFINEMENT OF PRH75 SUBCELLULAR LOCALIZATION. PRH75 IS
NUCLEOLAR PROTEIN 53 3.2.6. DETERMINATION OF THE NUCLEAR LOCALIZATION
SEQUENCE (NLS) OF PRH75 56 3.2.6.1. C-TERMINAL DELETIONS DELINEATED 81
AMINO ACIDS AT THE AMINO TERMINUS OF PRH75 CONTAINING A FUNCTIONAL NLS
56 3.2.6.2. NUCLEAR LOCALIZATION OF PRH75 IS MEDIATED BY AMINO ACIDS
59-81 57 3.2.7. BIOCHEMICAL ANALYSIS OF PRH75 61 3.2.7.1. OVEREXPRESSED
PRH75 DOES NOT EXHIBIT ATPASE ACTIVITY 61 3.2.7.2. PRH75 IS PRESENT IN A
LARGE PROTEIN COMPLEX OF ABOUT 500 KDA ... 61 3.2.8. FUNCTIONAL ANALYSIS
OF PRH75 67 III 3.2.8.2.2. DOMINANT-NEGATIVE MUTANTS OF PRH75 69 IV.
DISCUSSION 71 V. REFERENCES 81 VI. APPENDIX 98 6.1. MOLECULAR
CHARACTERIZATION OF PSBW, A NUCLEAR ENCODED COMPONENT OF PHOTOSYSTEM II
CORE COMPLEX FROM SPINACH [LORKOVIC ET AL., (1995) PROC. NATL. ACAD.
SCI. USA 92:8930-8934; LORKOVIC ET AL., (1995) IN P. MATIS (ED)
PHOTOSYNTHESIS: FROM LIGHT TO BIOSPHERE, P.743-746] 98 ACKNOWLEDGMENTS
104 PUBLICATIONS 105 CURRICULUM VITAE 106 IV
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| indexdate | 2024-07-09T18:15:19Z |
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| spelling | Lorković, Zdravko 1967- Verfasser (DE-588)120099640 aut Isolation and molecular characterization of cDNAs encoding PRH75, a nucleolar DEAD-box protein from higher plants von Zdravko Lorkovic 1997 V, 106 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Univ., Diss., 1997 Samenpflanzen (DE-588)4076988-4 gnd rswk-swf cDNS (DE-588)4212297-1 gnd rswk-swf Genklonierung (DE-588)4123275-6 gnd rswk-swf DEAD-box-Proteine (DE-588)4418310-0 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Samenpflanzen (DE-588)4076988-4 s DEAD-box-Proteine (DE-588)4418310-0 s cDNS (DE-588)4212297-1 s Genklonierung (DE-588)4123275-6 s DE-604 GBV Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=007936531&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Lorković, Zdravko 1967- Isolation and molecular characterization of cDNAs encoding PRH75, a nucleolar DEAD-box protein from higher plants Samenpflanzen (DE-588)4076988-4 gnd cDNS (DE-588)4212297-1 gnd Genklonierung (DE-588)4123275-6 gnd DEAD-box-Proteine (DE-588)4418310-0 gnd |
| subject_GND | (DE-588)4076988-4 (DE-588)4212297-1 (DE-588)4123275-6 (DE-588)4418310-0 (DE-588)4113937-9 |
| title | Isolation and molecular characterization of cDNAs encoding PRH75, a nucleolar DEAD-box protein from higher plants |
| title_auth | Isolation and molecular characterization of cDNAs encoding PRH75, a nucleolar DEAD-box protein from higher plants |
| title_exact_search | Isolation and molecular characterization of cDNAs encoding PRH75, a nucleolar DEAD-box protein from higher plants |
| title_full | Isolation and molecular characterization of cDNAs encoding PRH75, a nucleolar DEAD-box protein from higher plants von Zdravko Lorkovic |
| title_fullStr | Isolation and molecular characterization of cDNAs encoding PRH75, a nucleolar DEAD-box protein from higher plants von Zdravko Lorkovic |
| title_full_unstemmed | Isolation and molecular characterization of cDNAs encoding PRH75, a nucleolar DEAD-box protein from higher plants von Zdravko Lorkovic |
| title_short | Isolation and molecular characterization of cDNAs encoding PRH75, a nucleolar DEAD-box protein from higher plants |
| title_sort | isolation and molecular characterization of cdnas encoding prh75 a nucleolar dead box protein from higher plants |
| topic | Samenpflanzen (DE-588)4076988-4 gnd cDNS (DE-588)4212297-1 gnd Genklonierung (DE-588)4123275-6 gnd DEAD-box-Proteine (DE-588)4418310-0 gnd |
| topic_facet | Samenpflanzen cDNS Genklonierung DEAD-box-Proteine Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=007936531&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT lorkoviczdravko isolationandmolecularcharacterizationofcdnasencodingprh75anucleolardeadboxproteinfromhigherplants |