Import and function of inner envelope proteins from chloroplasts:
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2005
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | V, 91 Bl. Ill., graph. Darst. |
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| 245 | 1 | 0 | |a Import and function of inner envelope proteins from chloroplasts |c von Ahmed Mohamed Kamal Ahmed Nada |
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| 502 | |a München, Univ., Diss., 2005 | ||
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| adam_text | IMPORT AND FUNCTION OF INNER ENVELOPE PROTEINS FROM CHLOROPLASTS
DISSERTATION ZUR ERLANGUNG DES DOKTORGRADES DER FAKULTAT FIIR BIOLOGIE
DER LUDWIG-MAXIMILIANS-UNIVERSITAT MUNCHEN VORGELEGT VON AHMED MOHAMED
KAMAL AHMED NADA MUNCHEN 2005 CONTENTS CONTENTS ABBREVIATIONS AND
SYMBOLS IV I INTRODUCTION 1 II MATERIAL AND METHODS 9 II.L MATERIAL 9
II.L.L CHEMICALS . 9 II. II. II. II. II. .2 KITS 9 .3 ENZYMES 9 .4
MOLECULAR WEIGHT AND SIZE MARKERS 10 .5 DNA PRIMERS 10 1.6 BACTERIAL
STRAINS AND PLANT MATERIAL 10 II. 1.6.1 BACTERIAL STRAINS 10 II.L.6.2
PLANT MATERIAL 10 N. 1.7 VECTORS 11 II.1.8 GROWTH MEDIA 11 II. 1.9
RADIOISOTOPES 13 II.L.LOANTISERA 13 II.L.L 1 MEMBRANES 13 II. 1.12 FILMS
AND IMAGING PLATES 13 II.2 METHODS . . 14 11.2.1 GENERAL MOLECULAR
BIOLOGICAL METHODS 14 II.2.1.2PLASMID DNA ISOLATION 14
II.2.1.3POLYMERASE CHAIN REACTION (PCR) 14 11.2.1.4 CLONING TECHNIQUES
15 11.2.1.5 IN VITRO TRANSCRIPTION AND TRANSLATION 15 11.2.2 ISOLATION
OF CHLOROPLASTS 15 11.2.2.1 ISOLATION OF INTACT CHLOROPLASTS FROM PEA 15
11.2.2.2 ISOLATION OF INTACT CHLOROPLASTS FROM ARABIDOPSIS THALIANA 16
11.2.2.3 FRACTIONATION OF ISOLATED CHLOROPLASTS FROM ARABIDOPSIS
THALIANA 16 11.2.2.4 PREPARATION OF INNER ENVELOPE VESICLES 16 11.2.3
CHEMICAL TREATMENT OF CHLOROPLASTS BEFORE IMPORT 17 11.2.3.1 ATP
DEPLETION OF CHLOROPLASTS AND IN VITRO TRANSLATION PRODUCT 17 11.2.3.2
PROTEASE PRETREATMENT OF ISOLATED INTACT CHLOROPLASTS 17 II.2.3/3
CHLOROPLASTS PRETREATMENT WITH SPECIFIC CHEMICALS 17 11.2.4 IMPORT
EXPERIMENTS 18 11.2.4.1 IMPORT OF RADIOACTIVE-LABELLED PROTEINS INTO
INTACT CHLOROPLASTS 18 11.2.4.2 IMPORT OF RADIOACTIVELY LABELLED
PROTEINS INTO INNER ENVELOPE VESICLES 18 11.2.5 CHEMICAL TREATMENT OF
CHLOROPLASTS AFTER IMPORT 19 11.2.5.1 PROTEASE POST-TREATMENT OF INTACT
CHLOROPLASTS AND ENVELOPE MEMBRANES 19 11.2.5.2 SUBORGANELLAR
LOCALIZATION OF THE IMPORTED CONSTRUCTS 19 11.2.5.3 CHEMICAL
CROSSLINKING 20 11.2.6 PRODUCTION OF ANTISENSE MUTANT 20 II.2.6.1
PREPARATION OF THE ANTISENSE CONSTRUCT 20 112.6.2 PREPARATION OF
AGROBACTERIUM COMPETENT CELLS 21 11.2.6.3 TRANSFORMATION OF
AGROBACTERIUM 21 11.2.6.4 PLANT TRANSFORMATION USING INFILTRATION METHOD
22 11.2.6.5 SELECTION OF TRANSGENIC SEEDS 22 11.2.6.6 PREPARATION OF
SAMPLES FOR ELECTRON MICROSCOPE 23 11.2.7 OVEREXPRESSION OF RECOMBINANT
PROTEINS 24 II.2.7.1 CELL/VECTOR SYSTEMS 24 CONTENTS 11.2.1.2
SOLUBILIZATION OF 33-KDA OXYGEN EVOLVING COMPLEX SUBUNIT (POE33)
INCLUSION BODIES 24 11.2.7.3 OVEREXPRESSION USING IMPACT SYSTEM 25
11.2.7.4 OVEREXPRESSION USING RAPID TRANSLATION SYSTEM (RTS100) 25
11.2.8 METHODS FOR SEPARATION AND IDENTIFICATION OF PROTEINS 26 11.2.8.1
TOTAL PROTEIN EXTRACTION FROM PLANT TISSUE 26 11.2.8.2
SDS-POLYACRYLAMIDE-GEL-ELECTROPHORESIS (SDS-PAGE) 26 11.2.8.3 DETECTION
OF PROTEINS IN THE GELS 27 11.2.8.4 WESTERN TRANSFER 27 11.2.8.5
IMMUNODECORATION OF PROTEINS WITH ANTIBODIES 27 11.2.9 GENERAL METHODS
OF PROTEIN BIOCHEMISTRY 28 III RESULTS 29 111.1 CHARACTERIZATION OF
TIC32 IMPORT PATHWAY: 29 111.1.1 TIC32, A PROTEIN MEMBER OF THE TIC
COMPLEX, HAS NO TYPICAL PRESEQUENCE 29 111.1.2 IMPORT OF TIC32 INTO
CHLOROPLASTS 30 111.1.3 ENERGY REQUIREMENTS FOR IMPORT OF TIC32 INTO
CHLOROPLASTS 30 111.1.3.1 ATP DEPENDENCE 30 111.1.3.2 GTP DEPENDENCE 32
III. 1.3.3 PROTEASE SENSITIVITY TEST FOR TIC32 33 111.1.3.4 EXTRACTION
OF MEMBRANE INTEGRATED PROTEINS AFTER IMPORT WITH UREA 34 111.1.3.5
DEPLETION OF ATP FROM THE IMPORT REACTION 35 111.1.3.6 LOCALIZATION OF
TIC32 AFTER IMPORT 36 111.1.4 HOW IS TIC32 TARGETED TO CHLOROPLASTS? 37
III. 1.4 REQUIREMENT OF PROTEASE-SENSITIVE CHLOROPLASTIC SURFACE
PROTEINS 38 III. 1.5 TRANSLOCATION ACROSS THE OUTER ENVELOPE THROUGH
TOC75(III) 39 III. 1.5 INVOLVEMENT OF THE TIC COMPLEX DURING
TRANSLOCATION 42 111.1.6 INVESTIGATION OF PROTEIN PARTNERS INTERACTING
WITH TIC32 DURING IMPORT 43 111.1.7 IMPORT INTO INNER ENVELOPE MEMBRANE
VESICLES 46 111.2 CHARACTERIZATION OF A NEW ZINC PROTEASE 48 111.2.1
ANEW ZINC PROTEASE MIGHT WORK ON PROCESSING OF ITOC75 48 111.2.2
LOCALIZATION OF THE NEW ZINC PROTEASE (ARASP) 49 111.2.3 ARASP IS
ENCODED BY A LIGHT INDUCIBLE GENE 50 111.2.4 OVEREXPRESSION OF ARASP 51
111.2.4,1 OVEREXPRESSION OF ARASP USING THE IMPACT SYSTEM 51 111.2.4.2
CLEAVAGE OF ARASP FROM INTEIN TAG BY REDUCING REAGENT 52 111.2.4.3
OVEREXPRESSION OF THE ZINC PROTEASE (ARASP) USING THE RTS SYSTEM 54
111.2.5 ENZYME-SUBSTRATE RELATIONSHIP BETWEEN ARASP AND ITOC75MSSU 54
111.2.6 INHIBITION OF ARASP SYNTHESIZED USING RNA ANTISENSE TECHNIQUE 55
111.2.6.1 THE LEVEL OF ARASP PROTEIN WAS REDUCED IN THE TRANSGENIC
PLANTS: 55 111.2.6.2 GERMINATION RATE, GROWTH RATE AND PHENOTYPE OF THE
ANTISENSE PLANTS 56 111.2.6.3 INVESTIGATION OF CHLOROPLASTS OF THE WILD
TYPE AND THE ANTISENSE TRANSGENIC SEEDLINGS VIA ELECTRON MICROSCOPY 57
111.2.6.3 INVESTIGATION OF CHLOROPLASTS OF THE WILD TYPE AND THE
ANTISENSE TRANSGENIC SEEDLINGS VIA ELECTRON MICROSCOPY 58 111.2.6.4
INVESTIGATION OF ARASP SUBSTRATE USING WESTERN BLOT ANALYSIS FOR WILD
TYPE AND ANTISENSE PLANTS 58 IV DISCUSSION 61 IV.L. CHARACTERIZATION OF
TIC32 IMPORT PATHWAY 61 IV. 1.1 TIC32 IS A TIC SUBUNIT PROTEIN WITHOUT
CLEAVABLE TRANSIT PEPTIDE 61 IV.L.2 N-PROXIMAL AMINO ACIDS ARE ESSENTIAL
FOR TIC32 TARGETING AND IMPORT INTO CHLOROPLASTS 61 CONTENTS IV. 1.3
TIC32 IMPORTS INTO THE INNER ENVELOPE DIRECTLY WITHOUT ENTERING THE
STROMA 62 IV.1.4 STROMAL CHAPERONES ARE NOT INVOLVED IN THE TIC32 IMPORT
PROCESS 63 IV.1.5 TIC32 DOES NOT REQUIRE PROTEASE SENSITIVE RECEPTORS 63
IV.1.6 TOC75 IS NOT INVOLVED IN THE TIC32 IMPORT PATHWAY 64 . IV.1.7 TIC
COMPLEX IS NOT INVOLVED IN THE TIC32 IMPORT PATHWAY 65 IV. 1.8 TIC22 MAY
HAVE A ROLE DURING THE TIC32 IMPORT PATHWAY 65 IV.L.9 WHAT IS THE TIC32
IMPORT PATHWAY? 66 IV.1.10 WHICH INNER-ENVELOPE SORTING PATHWAY DOES
TIC32 USE? 67 IV.1.11 THE INSIGHT INTO THE FUTURE EXPERIMENTS 68 IV.2
CHARACTERIZATION OF A NEW ZINC PROTEASE 69 IV.2.1 THE NEW ZINC PROTEASE
MIGHT HAVE AN IMPORTANT ROLE IN THE IMPORT OF SOME PREPROTEINS INTO THE
CHLOROPLASTS 69 IV.2.2 ARASP IS A MEMBRANE PROTEASE PRESENT IN THE
CHLOROPLAST 69 IV.2.3 HETEROLOGOUS EXPRESSION OF ARASP 70 IV.2.4 ARASP
IS AN ESSENTIAL PROTEASE FOR PLANT SURVIVAL 71 IV.2.4 ARASP IS ESSENTIAL
PROTEASE FOR CHLOROPLAST BIOGENESIS 72 IV.2.5 SIGMA 2 MAY BE A SUBSTRATE
FOR ARASP 72 IV.2.6 THE INSIGHT INTO FUTURE EXPERIMENTS 74 V. SUMMARY 75
VI. ZUSAMMENFASSUNG 76 VII. REFERENCES ., 77 CURRICULUM VITAE 90
ACKNOWLEDGEMENT 91 IN
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| author | Nada, Ahmed Mohamed Kamal Ahmed 1972- |
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| language | English |
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| spelling | Nada, Ahmed Mohamed Kamal Ahmed 1972- Verfasser (DE-588)130082406 aut Import and function of inner envelope proteins from chloroplasts von Ahmed Mohamed Kamal Ahmed Nada 2005 V, 91 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Univ., Diss., 2005 Chloroplast (DE-588)4147821-6 gnd rswk-swf Hüllproteine (DE-588)4273116-1 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Chloroplast (DE-588)4147821-6 s Hüllproteine (DE-588)4273116-1 s b DE-604 GBV Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=013162061&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Nada, Ahmed Mohamed Kamal Ahmed 1972- Import and function of inner envelope proteins from chloroplasts Chloroplast (DE-588)4147821-6 gnd Hüllproteine (DE-588)4273116-1 gnd |
| subject_GND | (DE-588)4147821-6 (DE-588)4273116-1 (DE-588)4113937-9 |
| title | Import and function of inner envelope proteins from chloroplasts |
| title_auth | Import and function of inner envelope proteins from chloroplasts |
| title_exact_search | Import and function of inner envelope proteins from chloroplasts |
| title_full | Import and function of inner envelope proteins from chloroplasts von Ahmed Mohamed Kamal Ahmed Nada |
| title_fullStr | Import and function of inner envelope proteins from chloroplasts von Ahmed Mohamed Kamal Ahmed Nada |
| title_full_unstemmed | Import and function of inner envelope proteins from chloroplasts von Ahmed Mohamed Kamal Ahmed Nada |
| title_short | Import and function of inner envelope proteins from chloroplasts |
| title_sort | import and function of inner envelope proteins from chloroplasts |
| topic | Chloroplast (DE-588)4147821-6 gnd Hüllproteine (DE-588)4273116-1 gnd |
| topic_facet | Chloroplast Hüllproteine Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=013162061&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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