Genome sequencing of Thermoplasma acidophilum and characterization of the translation-associated ATPase of 43 kDa (TAA43), a member of the AAA+ family:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
2002
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | München, Techn. Univ., Diss., 2002 |
| Beschreibung: | V, 100 Bl. Ill., graph. Darst. |
Internformat
MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV014688815 | ||
| 003 | DE-604 | ||
| 005 | 20031103 | ||
| 007 | t| | ||
| 008 | 020829s2002 xx ad|| m||| 00||| eng d | ||
| 016 | 7 | |a 965259021 |2 DE-101 | |
| 035 | |a (OCoLC)216897291 | ||
| 035 | |a (DE-599)BVBBV014688815 | ||
| 040 | |a DE-604 |b ger |e rakwb | ||
| 041 | 0 | |a eng | |
| 049 | |a DE-91 |a DE-12 | ||
| 084 | |a BIO 180d |2 stub | ||
| 084 | |a BIO 310d |2 stub | ||
| 100 | 1 | |a Santos Martinez, Martha Leticia |d 1971- |e Verfasser |0 (DE-588)124043526 |4 aut | |
| 245 | 1 | 0 | |a Genome sequencing of Thermoplasma acidophilum and characterization of the translation-associated ATPase of 43 kDa (TAA43), a member of the AAA+ family |c Martha Leticia Santos Martinez |
| 264 | 1 | |c 2002 | |
| 300 | |a V, 100 Bl. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a München, Techn. Univ., Diss., 2002 | ||
| 650 | 0 | 7 | |a Genanalyse |0 (DE-588)4200230-8 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Thermoplasma acidophilum |0 (DE-588)4185162-6 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Thermoplasma acidophilum |0 (DE-588)4185162-6 |D s |
| 689 | 0 | 1 | |a Genanalyse |0 (DE-588)4200230-8 |D s |
| 689 | 0 | |5 DE-604 | |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009961611&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-009961611 | |
Datensatz im Suchindex
| _version_ | 1816444923559280640 |
|---|---|
| adam_text |
CONTENTS
I
CONTENTS
GENOME SEQUENCING OF
THERMOPLASMA ACIDOPHILUM AND
CHARACTERIZATION OF THE TRANSLATION-ASSOCIATED ATPASE OF 43 KDA
(TAA43), A MEMBER OF THE AAA+ FAMILY
1. SUMMARY 1
2. INTRODUCTION 3
2.1. BACKGROUND 3
2.1.1 .GENERAL FEATURES 3
2.1.2. DESCRIPTION OF ATPASES 5
2.1.3. AAA FAMILY 6
2.1.4. AAA FAMILY TREE 7
2.2. DESCRIPTION OF AAA SUBFAMILIES 8
2.2.1. MEIOSIS/MITOCHONDRIA 8
2.2.2. SECRETION/NEUROTRANSMISSION 9
2.2.3. PEROXISOMAL PROTEINS 9
2.2.4. SUBUNITS OF THE 26S PROTEASOME 10
2.2.5. CELL DIVISION CYCLE/CENTROSOME/ER HOMOTYPIC FUSION 10
2.2.6. PROTEASES 11
2.3. AIMS OF STUDY 12
3. MATERIALS AND METHODS 13
3.1 MATERIALS 13
3.1.1. CHEMICALS 13
3.1.2. BACTERIAL CULTURE MEDIA 14
3.1.3. ANTIBIOTICS 14
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/965259021
CONTENTS
_
JJ
3.1.4. KITS 14
3.1.5. ENZYMES AND ANTIBODIES 15
3.1.6. SIZE MARKERS 15
3.1.7. COLUMNS AND CHROMATOGRAPHIC MEDIA 16
3.1.8. BACTERIAL STRAINS 16
3.1.9. VECTORS 16
3.1.10. PRIMERS 17
3.2. METHODS 18
3.2.1. CELL CULTURE METHODS ' 18
3.2.1.1. GROWTH OF
T. ACIDOPHILUM 18
3.2.1.2. STORAGE OF
T. ACIDOPHILUM 19
3.2.1.3. GROWTH AND STORAGE OF
E. COLI CELLS 19
3.3. MOLECULAR BIOLOGY 20
3.3.1. BASIC TECHNIQUES - 20
3.3.1.1. PLASMID MINIPREP PROCEDURE 20
3.3.1.2. DETERMINATION OF DNA CONCENTRATION 20
3.3.1.3. AGAROSE GEL ELECTROPHORESIS 21
3.3.1.4. DNA SEQUENCING 21
3.3.1.5. POLYMERASE CHAIN REACTION 21
3.3.1.6. RESTRICTION ENDONUCLEASE DIGESTION 22
3.3.1.7. DEPHOSPHORYLATION 22
3.3.1.8. LIGATION 22
3.3.1.9. PREPARATION OF COMPETENT CELLS USING CACL 22
3.3.1.10. TRANSFORMATION OF E. COLI CELLS 22
3.3.1.11. MUTAGENESIS 23
3.3.2. GENOME SEQUENCING . 24
3.3.2.1. ISOLATION OF CHROMOSOMAL DNA FROM
T. ACIDOPHILUM 24
3.3.2.2. PHAGEMID LIBRARIES 25
3.3.2.3. COSMID LIBRARIES 25
3.3.2.4. LONG RANGE PCR 25
3.3.2.5. SEQUENCING THE REVERSE STRAND OF COSMIDS / 25
3.3.2.6. ASSEMBLY AND EDITING 26
3.4. PROTEIN CHEMICAL METHODS 27
3.4.1. EXPRESSION AND PURIFICATION OF RECOMBINANT
HIS-TAGGED PROTEIN UNDER NATIVE CONDITIONS 27
3.4.2. DETERMINATION OF PROTEIN CONCENTRATION 27
3.4.3. DENATURING SDS-PAGE 27
3.4.4. NATIVE PAGE 28
3.4.5. COLLOIDAL COOMASSIE BRILLIANT BLUE G-250 STAINING 29
3.4.6. PREPARATION OF ANTISERUM 29
CONTENTS
M
3.4.7. PROTEIN TRANSFER ONTO NITROCELLULOSE 29
3.4.8. IMMUNOBLOTTING WITH ALKALINE PHOPHATASE 30
3.4.9. GRADIENT ULTRACENTRIFUGATION 31
3.4.10. SIZE EXCLUSION CHROMATOGRAPHY 31
3.4.11. CHARACTERIZATION OF ATPASE ACTIVITY 31
3.5. COMPUTER ANALYSIS 33
3.5.1. PHYLOGENETIC TREE CLASSIFICATION OF TAA43 AND SEQUENCE ANALYSIS
33
3.5.2. DNA AND PROTEIN SEQUENCE ANALYSIS , 33
3.6. FUNCTIONAL STUDIES 34
3.6.1. FOLDASE/UNFOLDASE STUDIES 34
3.6.2. ELECTROPHORETIC MOBILITY SHIFT ASSAY 35
3.6.2.1. LABELLING OF OLIGONUCLEOTIDES AT THE 3 'END 35
3.6.2.2. GEL SHIFT REACTION AND CHEMILUMINESCENCE DETECTION 35
3.6.3. IMMUNODEPLETION ASSAYS 36
3.6.4. DETERMINATION OF PEPTIDASE ASSAY ~ 36
3.6.5. CASEIN DEGRADATION ASSAY 36
3.6.6. TWO-HYBRID SYSTEM IN BACTERIA 37
3.6.6.1. PLASMID LIBRARY CONSTRUCTION INTO PUTL8 37
3.6.6.2. CLONING TAA43 INTO PT25 37
3.6.6.3. PREPARATION OF BTH101 COMPETENT CELLS 37
3.6.6.4. CO-EXPRESSION AND SELECTION OF INTERACTING
HYBRID PROTEINS 37
3.6.6.5.13-GALACTOSIDASE ASSAY 38
3.6.7. CO-PRECIPITATION STUDIES 38
3.6.8. PROTEIN SEQUENCING ANALYSIS 39
3.6.9. COMPUTER ANALYSIS 39
4. RESULTS ' 40
4.1
. T. ACIDOPHILUM GENOME PROJECT 40
4.2. TAA43 CHARACTERIZATION 41
4.2.1. SEQUENCE ANALYSIS OF THE TAA43 41
4.2.2. PHYLOGENETIC TREE CLASSIFICATION OF TAA43 WITHIN THE AAA FAMILY
43
4.2.3. SEQUENCE ANALYSIS OF THE N-TERMINAL DOMAIN OF TAA43 45
4.2.4. CLONING 45
4.2.5. EXPRESSION AND PURIFICATION OF RECOMBINANT TAA43
PROTEIN IN E. COLI 47
4.2.6. DETECTION OF IN VIVO EXPRESSION IN
T. ACIDOPHILUM 48
4.2.7. DETERMINATION OF RELATIVE MOLECULAR MASS 49
4.2.8. ENZYMATIC CHARACTERIZATION 52
4.2.9. KINETICS AND INHIBITION ANALYSIS 54
4.2.10. SITE-DIRECTED MUTAGENESIS OF CONSERVED RESIDUES 55
CONTENTS
_/
4.3. FUNCTIONAL STUDIES 57
4.3.1.
T. ACIDOPHILUM GENOME ANALYSIS - THE TRANSLATION-ASSOCIATED
ATPASE (TAA43) GENE AND ITS NEIGHBOURS IN THE GENOME- 57
4.3.2. CHAPERONIN-ASSISTED PROTEIN FOLDING OF THE ENZYME
RHODANESE BY TAA43 59
4.3.3. PROTEOLYTIC DEGRADATION PATHWAY "VIA PROTEASOME" 61
4.3.4. DNA-BINDING ASSAY 63
4.3.5.
IN VIVO ANALYSIS OF PROTEIN-PROTEIN INTERACTIONS , 65
4.3.5.1. PREY LIBRARY CONSTRUCTION WITH PUT18 66
4.3.5.2. BAIT CLONING:
TAA43 GENE INTO PT25 66
4.3.5.3. SCREENING OF POSITIVE INTERACTING COLONIES BY THE
TWO-HYBRID SYSTEM 67
4.3.5.4. POSITIVE INTERACTING COLONIES 68
4.3.6.
IN VITRO ANALYSIS OF PROTEIN-PROTEIN INTERACTIONS 70
5. DISCUSSION
YY
72
5.1. BIOCHEMICAL CHARACTERIZATION OF TAA43 72
5.1.1. CONFORMATIONAL STATE 72
5.1.2. ATPASE ACTIVITY 73
5.1.3. MUTATIONAL ANALYSIS 74
5.2. FUNCTIONAL ANALYSIS 75
5.2.1. TWO-HYBRID SYSTEM IN BACTERIA 75
5.2.2. CO-PRECIPITATION ANALYSIS 76
*
5.3. DESCRIPTION OF THE TAA43 INTERACTING PROTEINS 78
5.3.1. RIBOSOMES 78
5.3.2. RIBONUCLEOTIDE REDUCTASE 79
5.3.3. INOSINE-5'-MONOPHOSPHATE DEHYDROGENASE 81
5.3.4. AMINOACYL-T-RNA SYNTHETHASES ' 81
5.3.5. TRANSLATION INITIATION FACTOR 82
5.3.6. TRANSLATION ELONGATION FACTOR 82
5.3.7. IRON-DEPENDENT TRANSCRIPTION REPRESSOR AND FERRIC
TRANSPORT ATP-BINDING PROTEIN AFUC 82
5.3.8. CELL DIVISION CONTROL 83
5.3.9. POLYKETIDE SYNTHASE 83
5.4. HYPOTHESIS FOR THE CELLULAR ROLE OF TAA43 84
5.5. FUTURE WORK
86
CONTENTS
_
6. BIBLIOGRAPHY 87
7. ABBREVIATIONS 98
8. ACKNOWLEDGMENTS 100 |
| any_adam_object | 1 |
| author | Santos Martinez, Martha Leticia 1971- |
| author_GND | (DE-588)124043526 |
| author_facet | Santos Martinez, Martha Leticia 1971- |
| author_role | aut |
| author_sort | Santos Martinez, Martha Leticia 1971- |
| author_variant | m m l s mml mmls |
| building | Verbundindex |
| bvnumber | BV014688815 |
| classification_tum | BIO 180d BIO 310d |
| ctrlnum | (OCoLC)216897291 (DE-599)BVBBV014688815 |
| discipline | Biologie |
| format | Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>00000nam a2200000 c 4500</leader><controlfield tag="001">BV014688815</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">20031103</controlfield><controlfield tag="007">t|</controlfield><controlfield tag="008">020829s2002 xx ad|| m||| 00||| eng d</controlfield><datafield tag="016" ind1="7" ind2=" "><subfield code="a">965259021</subfield><subfield code="2">DE-101</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)216897291</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)BVBBV014688815</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-91</subfield><subfield code="a">DE-12</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">BIO 180d</subfield><subfield code="2">stub</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">BIO 310d</subfield><subfield code="2">stub</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Santos Martinez, Martha Leticia</subfield><subfield code="d">1971-</subfield><subfield code="e">Verfasser</subfield><subfield code="0">(DE-588)124043526</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Genome sequencing of Thermoplasma acidophilum and characterization of the translation-associated ATPase of 43 kDa (TAA43), a member of the AAA+ family</subfield><subfield code="c">Martha Leticia Santos Martinez</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2002</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">V, 100 Bl.</subfield><subfield code="b">Ill., graph. Darst.</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">n</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">nc</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="500" ind1=" " ind2=" "><subfield code="a">München, Techn. Univ., Diss., 2002</subfield></datafield><datafield tag="650" ind1="0" ind2="7"><subfield code="a">Genanalyse</subfield><subfield code="0">(DE-588)4200230-8</subfield><subfield code="2">gnd</subfield><subfield code="9">rswk-swf</subfield></datafield><datafield tag="650" ind1="0" ind2="7"><subfield code="a">Thermoplasma acidophilum</subfield><subfield code="0">(DE-588)4185162-6</subfield><subfield code="2">gnd</subfield><subfield code="9">rswk-swf</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4113937-9</subfield><subfield code="a">Hochschulschrift</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="689" ind1="0" ind2="0"><subfield code="a">Thermoplasma acidophilum</subfield><subfield code="0">(DE-588)4185162-6</subfield><subfield code="D">s</subfield></datafield><datafield tag="689" ind1="0" ind2="1"><subfield code="a">Genanalyse</subfield><subfield code="0">(DE-588)4200230-8</subfield><subfield code="D">s</subfield></datafield><datafield tag="689" ind1="0" ind2=" "><subfield code="5">DE-604</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">DNB Datenaustausch</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009961611&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="943" ind1="1" ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-009961611</subfield></datafield></record></collection> |
| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV014688815 |
| illustrated | Illustrated |
| indexdate | 2024-11-22T17:34:28Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-009961611 |
| oclc_num | 216897291 |
| open_access_boolean | |
| owner | DE-91 DE-BY-TUM DE-12 |
| owner_facet | DE-91 DE-BY-TUM DE-12 |
| physical | V, 100 Bl. Ill., graph. Darst. |
| publishDate | 2002 |
| publishDateSearch | 2002 |
| publishDateSort | 2002 |
| record_format | marc |
| spelling | Santos Martinez, Martha Leticia 1971- Verfasser (DE-588)124043526 aut Genome sequencing of Thermoplasma acidophilum and characterization of the translation-associated ATPase of 43 kDa (TAA43), a member of the AAA+ family Martha Leticia Santos Martinez 2002 V, 100 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Techn. Univ., Diss., 2002 Genanalyse (DE-588)4200230-8 gnd rswk-swf Thermoplasma acidophilum (DE-588)4185162-6 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Thermoplasma acidophilum (DE-588)4185162-6 s Genanalyse (DE-588)4200230-8 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009961611&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Santos Martinez, Martha Leticia 1971- Genome sequencing of Thermoplasma acidophilum and characterization of the translation-associated ATPase of 43 kDa (TAA43), a member of the AAA+ family Genanalyse (DE-588)4200230-8 gnd Thermoplasma acidophilum (DE-588)4185162-6 gnd |
| subject_GND | (DE-588)4200230-8 (DE-588)4185162-6 (DE-588)4113937-9 |
| title | Genome sequencing of Thermoplasma acidophilum and characterization of the translation-associated ATPase of 43 kDa (TAA43), a member of the AAA+ family |
| title_auth | Genome sequencing of Thermoplasma acidophilum and characterization of the translation-associated ATPase of 43 kDa (TAA43), a member of the AAA+ family |
| title_exact_search | Genome sequencing of Thermoplasma acidophilum and characterization of the translation-associated ATPase of 43 kDa (TAA43), a member of the AAA+ family |
| title_full | Genome sequencing of Thermoplasma acidophilum and characterization of the translation-associated ATPase of 43 kDa (TAA43), a member of the AAA+ family Martha Leticia Santos Martinez |
| title_fullStr | Genome sequencing of Thermoplasma acidophilum and characterization of the translation-associated ATPase of 43 kDa (TAA43), a member of the AAA+ family Martha Leticia Santos Martinez |
| title_full_unstemmed | Genome sequencing of Thermoplasma acidophilum and characterization of the translation-associated ATPase of 43 kDa (TAA43), a member of the AAA+ family Martha Leticia Santos Martinez |
| title_short | Genome sequencing of Thermoplasma acidophilum and characterization of the translation-associated ATPase of 43 kDa (TAA43), a member of the AAA+ family |
| title_sort | genome sequencing of thermoplasma acidophilum and characterization of the translation associated atpase of 43 kda taa43 a member of the aaa family |
| topic | Genanalyse (DE-588)4200230-8 gnd Thermoplasma acidophilum (DE-588)4185162-6 gnd |
| topic_facet | Genanalyse Thermoplasma acidophilum Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009961611&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT santosmartinezmarthaleticia genomesequencingofthermoplasmaacidophilumandcharacterizationofthetranslationassociatedatpaseof43kdataa43amemberoftheaaafamily |