Homo-phytochelatin biosynthesis and cobalt detoxification in plants:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2001
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | VIII, 153 S. Ill., graph. Darst. |
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Datensatz im Suchindex
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| adam_text | DISSERTATION ZUR ERLANGUNG DES DOKTORGRADES DER FAKULTAT FIIR CHEMIE UND
PHARMAZIE DER LUDWIG-MAXIMILIANS-UNIVERSITAT MUNCHEN HOMO-PHYTOCHELATIN
BIOSYNTHESIS AND COBALT DETOXIFICATION IN PLANTS MATJAZ OVEN AUS
LJUBLJANA / SLOWENIEN 2001 CONTENTS I CONTENTS A. INTRODUCTION 1 B.
MATERIALS AND METHODS 21 1. ORGANISMS 21 1.1. PLANT CELL CULTURES 21
1.2. MICROORGANISMS AND COMPOSITION OF MEDIA 21 2. CHEMICALS 23 3.
EQUIPMENT ,. 23 4. DETERMINATION OF ENZYMATIC ACTIVITY 24 4.1.
PHYTOCHELATIN SYNTHASE ASSAY 24 4.2. CITRATE LYASE ASSAY 25 5.
ANALYTICAL METHODS 26 5.1. DETERMINATION OF COBALT 26 5.2. DETERMINATION
OF PROTEINS 27 5.3. DETERMINATION OF NUCLEIC ACIDS 27 5.4. DETERMINATION
OF CYSTEINE BY GAITONDE CYSTEINE ASSAY 27 6. CHROMATOGRAPHY 28 6.1.
THIN-LAYER CHROMATOGRAPHY (TLC) 28 6.2. SIZE-EXCLUSION CHROMATOGRAPHY 29
6.3. HPLC METHODS 29 6.3.1. EXTRACTION METHOD AND SAMPLE PREPARATION 30
6.3.2. ANALYTICAL HPLC WITH DTNB POST-COLUMN DERIVATIZATION 31 6.3.3.
ANALYTICAL HPLC WITHOUT POSTCOLUMN DERIVATIZATION 32 7. MASS
SPECTROMETRY 33 8. ENZYMES, KITS AND SPECIAL CHEMICALS USED IN MOLECULAR
BIOLOGY PROCEDURES 33 8.1. COMMONLY USED ENZYMES IN MOLECULAR BIOLOGY
PROCEDURES 33 8.2. MOLECULAR BIOLOGY KITS 34 8.3. SPECIAL CHEMICALS USED
IN THE MOLECULAR BIOLOGY PROCEDURES 34 9. NUCLEIC ACIDS 35 9.1. DNA 35
9.2. ISOLATION OF NUCLEIC ACIDS 36 9.2.1. ISOLATION OF TOTAL RNA 36
9.2.2. ISOLATION OF POLY A + RNA(MRNA) 38 0 CONTENTS 9.2.3. ISOLATION OF
GENOMICDNA 38 9.2.4. ISOLATION OF PLASMID DNA FROM BACTERIA 39 9.2.5.
DNA ISOLATION FROM AGAROSE GELS 40 9.3. ENZYMATIC MODIFICATIONS OF
NUCLEIC ACIDS 40 9.3.1. RESTRICTION ANALYSIS 40 9.3.2. LIGATIONS 41
9.3.3. REVERSE TRANSCRIPTION (CDNA SYNTHESIS) 42 10. GEL ELECTROPHORESIS
42 10.1. AGAROSE GEL ELECTROPHORESIS OF DNA 42 10.2. AGAROSE GEL
ELECTROPHORESIS OF RNA 43 10.3. SODIUM DODECYLSULPHATE - POLYACRYLAMIDE
GEL ELECTROPHORESIS (SDS-PAGE) 44 10.4. PROTEIN GEL DYING 46 11.
POLYMERASE CHAIN REACTION (PCR) 46 12. RADIOACTIVE LABELING OF NUCLEIC
ACIDS 47 13. BLOTTING TECHNIQUES 48 13.1. NORTHERN BLOT 48 13.2.
SOUTHERN BLOT 49 14. HYBRIDIZATION TECHNIQUES 50 15. DNA SEQUENCING 51
16. PREPARATION AND TRANSFORMATION OF COMPETENT E. COLI CELLS 52 16.1.
PREPARATION OF COMPETENT E. COLI CELLS . 52 16.2. TRANSFORMATION OF
COMPETENT E. COLI CELLS 53 17. ESTABLISHMENT OF CDNA LIBRARY IN X-PHAGES
53 17.1. CONSTRUCTION OF X ZAP-CDNA GIGAPACK III LIBRARY 53 17.2.
SCREENING OF A X ZAP-CDNA GIGAPACK III LIBRARY 55 17.3. IN VIVO EXCISION
57 18. EXPRESSION AND PURIFICATION OF RECOMBINANT PROTEINS 58 19.
COMPARISON OF GENETIC DATABASES 60 C. RESULTS 61 1. INVESTIGATION OF
COBALT COMPLEXATION IN PLANT CELL CULTURES 61 1.1. STUDIES OF COBALT
UPTAKE AND TOLERANCE BY C. COBALTICOLA SUSPENSION CELLS 61 1.1.1.
SUSCEPTIBILITY OF C. COBALTICOLA SUSPENSION CELLS TOWARDS COBALT 61
1.1.2. COBALT UPTAKE BY C. COBALTICOLA SUSPENSION CELLS 63 CONTENTS FFL^
1.1.3. KINETICS OF COBALT UPTAKE BY C. COBALTICOLA SUSPENSION CELLS 63
1.1.4. GROWTH SUPPRESSION OF C. COBALTICOLA SUSPENSION CELLS BY COBALT,
NICKEL, CADMIUM, AND COPPER 65 1.2. IDENTIFICATION OF COBALT-INDUCED
COMPOUNDS IN C. COBALTICOLA SUSPENSION CELLS 66 1.2.1. ARE
PHYTOCHELATINS INVOLVED IN COBALT COMPLEXATION? 66 1.2.2. INDUCTION OF
ORGANIC ACIDS 68 1.2.3. INDUCTION OF AMINO ACIDS 70 1.3. ROLE OF
CYSTEINE IN COBALT COMPLEXATION 72 2. HOMO-PHYTOCHELATIN SYNTHASE FROM
SOYBEAN (GLYCINE MAX): MOLECULAR CLONING, HETEROLOGOUS EXPRESSION,
ENZYME PURIFICATION AND CHARACTERIZATION 76 2.1. ISOLATION OF
PHYTOCHELATIN SYNTHASE-HOMOLOGOUS MRNA FRAGMENTS 78 2.1.1. DESIGN OF
DEGENERATED PCR PRIMERS 78 2.1.2. G. MAX CDNA SYNTHESIS AND PCR
PROCEDURE 78 2.1.3. SUBCLONING AND SEQUENCE ANALYSIS OF AMPLIFIED PCR
FRAGMENTS 80 2.2. ISOLATION OF FULL-LENGTH HOMO-PHYTOCHELATIN SYNTHASE
CDNA FROM G. MAX 81 2.2.1. G.MAX CDNA-LIBRARY SCREENING 82 2.2.4.
HOMOLOGY COMPARISON WITH GENETIC DATABASES 84 2.3. HETEROLOGOUS
EXPRESSION OF HOMO-PHYTOCHELATIN SYNTHASE FROM GLYCINE MAX AND
PHYTOCHELATIN SYNTHASE FROM ARABIDOPSIS THALIANA IN E. COLI 87 2.3.1.
CONSTRUCTION OF EXPRESSION VECTORS 88 2.3.2. IDENTIFICATION OF REACTION
PRODUCTS 90 2.3. SOUTHERN BLOT ANALYSIS 93 2.4. NORTHERN BLOT ANALYSIS
95 2.5. PURIFICATION OF RECOMBINANT HOMO-PHYTOCHELATIN SYNTHASE FROM
GLYCINE MAX AND RECOMBINANT PHYTOCHELATIN SYNTHASE FROM ARABIDOPSIS
THALIANA 97 2.6. CHARACTERIZATION OF HOMO-PHYTOCHELATIN SYNTHASE FROM
GLYCINE MAX AND PHYTOCHELATIN SYNTHASE FROM ARABIDOPSIS THALIANA 100
2.6.1. SYNTHESIS OF HOMO-PHYTOCHELATINS FROM HOMOGLUTATHIONE 100 2.6.2.
SYNTHESIS OF HOMO-PHYTOCHELATINS FROM GLUTATHIONE AND HOMOGLUTATHIONE
102 2.6.3. EFFECT OF GSH-CONCENTRATION ON HOMO-PHYTOCHELATIN SYNTHESIS
108 2.6.4. TEMPERATURE OPTIMUM FOR THE SYNTHESIS OF PHYTOCHELATINS AND
HOMO- PHYTOCHELATINS 109 2.6.5. PH OPTIMUM FOR THE SYNTHESIS OF
PHYTOCHELATINS AND HOMO-PHYTOCHELATINS 110 IV CONTENTS 2.6.6. KM VALUES
FOR SYNTHESIS OF PHYTOCHELATINS ILL 2.6.7. METAL ACTIVATION OF G. MAX
HOMO-PHYTOCHELATIN SYNTHASE AND A. THALIANA PC- SYNTHASE 112 2.7.
INVESTIGATIONS ON THE MECHANISM OF PHYTOCHELATIN SYNTHESIS 115 2.7.1.
ROLE OF BUFFERS IN PC-SYNTHASE ASSAY 115 2.7.2. ROLE OF (GS) 2 CD IN THE
SYNTHESIS OF PHYTOCHELATINS 117 2.7.3. INVESTIGATIONSION THE ACTIVATION
OF PHYTOCHELATIN SYNTHASES 119 D. DISCUSSION 123 E. SUMMARY 136 F.
REFERENCES ; 140
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| any_adam_object | 1 |
| author | Oven, Matjaž 1968- |
| author_GND | (DE-588)123277620 |
| author_facet | Oven, Matjaž 1968- |
| author_role | aut |
| author_sort | Oven, Matjaž 1968- |
| author_variant | m o mo |
| building | Verbundindex |
| bvnumber | BV013946674 |
| ctrlnum | (OCoLC)76296211 (DE-599)BVBBV013946674 |
| discipline | Medizin |
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| indexdate | 2024-07-09T18:54:52Z |
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| language | English |
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| physical | VIII, 153 S. Ill., graph. Darst. |
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| spelling | Oven, Matjaž 1968- Verfasser (DE-588)123277620 aut Homo-phytochelatin biosynthesis and cobalt detoxification in plants von Matjaž Oven 2001 VIII, 153 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Univ., Diss., 2001 Phytochelatine (DE-588)4305646-5 gnd rswk-swf Entgiftung (DE-588)4014873-7 gnd rswk-swf Cobalt (DE-588)4070047-1 gnd rswk-swf Pflanzen (DE-588)4045539-7 gnd rswk-swf Biosynthese (DE-588)4006902-3 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Pflanzen (DE-588)4045539-7 s Phytochelatine (DE-588)4305646-5 s Biosynthese (DE-588)4006902-3 s DE-604 Cobalt (DE-588)4070047-1 s Entgiftung (DE-588)4014873-7 s GBV Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009543329&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Oven, Matjaž 1968- Homo-phytochelatin biosynthesis and cobalt detoxification in plants Phytochelatine (DE-588)4305646-5 gnd Entgiftung (DE-588)4014873-7 gnd Cobalt (DE-588)4070047-1 gnd Pflanzen (DE-588)4045539-7 gnd Biosynthese (DE-588)4006902-3 gnd |
| subject_GND | (DE-588)4305646-5 (DE-588)4014873-7 (DE-588)4070047-1 (DE-588)4045539-7 (DE-588)4006902-3 (DE-588)4113937-9 |
| title | Homo-phytochelatin biosynthesis and cobalt detoxification in plants |
| title_auth | Homo-phytochelatin biosynthesis and cobalt detoxification in plants |
| title_exact_search | Homo-phytochelatin biosynthesis and cobalt detoxification in plants |
| title_full | Homo-phytochelatin biosynthesis and cobalt detoxification in plants von Matjaž Oven |
| title_fullStr | Homo-phytochelatin biosynthesis and cobalt detoxification in plants von Matjaž Oven |
| title_full_unstemmed | Homo-phytochelatin biosynthesis and cobalt detoxification in plants von Matjaž Oven |
| title_short | Homo-phytochelatin biosynthesis and cobalt detoxification in plants |
| title_sort | homo phytochelatin biosynthesis and cobalt detoxification in plants |
| topic | Phytochelatine (DE-588)4305646-5 gnd Entgiftung (DE-588)4014873-7 gnd Cobalt (DE-588)4070047-1 gnd Pflanzen (DE-588)4045539-7 gnd Biosynthese (DE-588)4006902-3 gnd |
| topic_facet | Phytochelatine Entgiftung Cobalt Pflanzen Biosynthese Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009543329&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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