Strongyloides ratti: identification, isolation and characterisation of Heat Shock Protein 10 and Heat Shock Protein 60
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Giessen
VVB Laufersweiler
2009
|
| Ausgabe: | 1.Aufl. |
| Schriftenreihe: | Edition scientifique
|
| Schlagworte: | |
| Online-Zugang: | Abstract Inhaltsverzeichnis |
| Beschreibung: | 123 S. Ill., graph. Darst. |
| ISBN: | 9783835954717 3835954717 |
Internformat
MARC
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|---|---|---|---|
| 001 | BV026885334 | ||
| 003 | DE-604 | ||
| 005 | 20110228 | ||
| 007 | t | ||
| 008 | 110326s2009 ad|| m||| 00||| eng d | ||
| 015 | |a 09N501307 |2 dnb | ||
| 020 | |a 9783835954717 |9 978-3-8359-5471-7 | ||
| 020 | |a 3835954717 |9 3-8359-5471-7 | ||
| 024 | 7 | |a urn:nbn:de:hebis:26-opus-71892 |2 urn | |
| 035 | |a (OCoLC)560430219 | ||
| 035 | |a (DE-599)DNB99845494X | ||
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| 082 | 0 | |a 614.5934 |2 22 | |
| 100 | 1 | |a Tazir, Yasmina |e Verfasser |0 (DE-588)139301178 |4 aut | |
| 245 | 1 | 0 | |a Strongyloides ratti |b identification, isolation and characterisation of Heat Shock Protein 10 and Heat Shock Protein 60 |c eingereicht von Yasmina Tazir |
| 250 | |a 1.Aufl. | ||
| 264 | 1 | |a Giessen |b VVB Laufersweiler |c 2009 | |
| 300 | |a 123 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 490 | 0 | |a Edition scientifique | |
| 502 | |a Zugl.: Giessen, Univ., Diss., 2009 | ||
| 650 | 7 | |a antibodies |2 cabt | |
| 650 | 7 | |a parasites |2 cabt | |
| 650 | 7 | |a nematoda |2 cabt | |
| 650 | 7 | |a strongyloididae |2 cabt | |
| 650 | 7 | |a helminths |2 cabt | |
| 650 | 7 | |a strongyloides ratti |2 cabt | |
| 650 | 7 | |a strongyloides stercoralis |2 cabt | |
| 650 | 7 | |a heat shock proteins |2 cabt | |
| 650 | 7 | |a immune response |2 cabt | |
| 650 | 7 | |a dna cloning |2 cabt | |
| 650 | 7 | |a transcription |2 cabt | |
| 650 | 7 | |a genome analysis |2 cabt | |
| 650 | 7 | |a open reading frames |2 cabt | |
| 650 | 7 | |a amino acid sequences |2 cabt | |
| 650 | 7 | |a host parasite relationships |2 cabt | |
| 650 | 7 | |a genes |2 cabt | |
| 650 | 7 | |a immunity |2 cabt | |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 856 | 4 | |u http://library.vetmed.fu-berlin.de/ResourceList/details/207755 |z lizenzfrei |3 Abstract | |
| 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=022411192&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 999 | |a oai:aleph.bib-bvb.de:BVB01-022411192 | ||
Datensatz im Suchindex
| _version_ | 1804145522440667136 |
|---|---|
| adam_text | IMAGE 1
TABLE OF CONTENTS
TABLE OF CONTENTS
I. TABLE OF CONTENTS
II. LIST OF FIGURES 6
III. LIST OF TABLES 8
1 INTRODUCTION 9
1.1 INTESTINAL PARASITES 9
1.2 STRONGYLOIDES RATTI 13
13 HEAT SHOCK PROTEINS 15
1.3.1 HEAT SHOCK PROTEIN 60 16
1.3.2 HEAT SHOCK PROTEIN 10 17
1.4 OBJECTIVE OF THIS THESIS 18
2 MATERIALS AND METHODS 19
2.1 REAGENTS, DISPOSALS, INSTRUMENTS 19
2.1.1 SOLUTIONS AND BUFFERS 19
2.1.2 MEDIA AND ADDITIVES 22
2.1.3 ENZYMES 23
2.1.4 MOLECULAR WEIGHT STANDARDS 24
2.1.5 PRIMER 24
2.1.6 PLASMIDS 27
2.1.7 BACTERIA AND YEAST STRAINS 35
2.1.8 ANTIBODIES 35
2.1.9 COMPUTER-BASED SEQUENCE ANALYSIS 36
2.2 STRONGYLOIDES RATTI LIFE CYCLE AND CULTURING 37
2.2.1 ANIMALS 37
2.2.2 HOST INFECTION 37
2.2.3 HOST IMMUNISATION 37
2.2.4 BAERMANN TECHNIQUE 37
23 MOLECULAR BIOLOGICAL METHODS _ 38
2.3.1 PRODUCTION OF COMPETENT BACTERIA (NISHIMURA ET AL 1990) 38
2.3.2 TRANSFORMATION OF DNA INTO BACTERIA 39
2.3.3 PURIFICATION OF PLASMID DNA 39
BIBLIOGRAFISCHE INFORMATIONEN HTTP://D-NB.INFO/99845494X
DIGITALISIERT DURCH
IMAGE 2
TABLE OF CONTENTS
2.3.3.1 PLASMID PURIFICATION FROM BACTERIAL CULTURES 39
2.3.3.1.1 MINI-SCALE PLASMID ISOLATION 39
2.3.3.1.2 ALTERNATIVE MINI-SCALE PLASMID ISOLATION 39
2.3.3.1.3 PLASMID-DNA ISOLATION FROM 100 ML CULTURES (MIDI PREPS) 40
2.3.3.1.4 DNA-FRAGMENT PURIFICATION 40
2.3.3. 1.5 DNA-FRAGMENT EXTRACTION FROM AGAROSE GELS 40
2.3.4 TOTAL RNA ISOLATION FROM S. RATTI INFECTIOUS LARVAE OR PARASITIC
FEMALES 40
2.3.4.1 TOTAL RNA ISOLATION 40
2.3.4.2 PHENOL/CHLOROFORM EXTRACTION 41
2.3.4.3 PRECIPITATION OF RNA 41
2.3.5 GDNA ISOLATION FROM S. RATTI INFECTIOUS LARVAE AND PRECIPITATION
42
2.3.6 POLYMERASE CHAIN REACTION (PCR) 42
2.3.7 5 AND 3 CDNAS AMPLIFICATION 42
2.3.8 DNA AGAROSE GEL ELECTROPHORESIS 43
2.3.9 SEQUENCING OF DNA 43
2.3.10 ENZYMATIC MANIPULATION OF DNA 43
2.3.10.1 RESTRICTION ANALYSIS 43
2.3.10.2 PLASMID DNA FRAGMENT 44
2.3.10.3 ENZYMATIC MANIPULATION OF VECTOR DNA PRIOR TO LIGATION 44
2.3.10.4 LIGATION OF PLASMID VECTOR AND INSERT DNA 44
2.3.11 PHOTOMETRIC QUANTIFICATION OF NUCLEIC ACIDS 45
2.3.12 SOUTHERN BLOT ANALYSIS 45
2.3.12.1 RANDOM PRIME RNA LABELLING 46
2.3.13 WHOLE MOUNT IN SITU HYBRIDISATION 46
2.3.13.1 TISSUE PREPARATION 46
2.3.13.1.1 PRIMARY FIXATION OF IL3 46
2.3.13.1.2 FIXATION OF IL3 ONTOSLIDES 47
2.3.13.1.3 KRYO-BLOCK PREPARATION 47
2.3.13.2 DIG-LABELLING OF RNA PROBES 48
2.3.13.3 PREHYBRIDISATION, HYBRIDISATION AND POSTHYBRIDISATION
PROCEDURES 48
2.3.13.4 DETECTION PROCEDURE 49
2.4 CELL CULTURES . 49
2.4.1 YEAST CELL CULTURE 49
2.5 PROTEIN BIOCHEMICAL METHODS 50
2.5.1 MAMMALIAN TWO-HYBRID 50
IMAGE 3
TABLE OF CONTENTS
2.5.1.1 TRANSFECTION 52
2.5.1.2 RENILLA LUCIFERASE ASSAY 53
2.5.2 YEAST TWO-HYBRID 53
2.5.2.1 FILTER AND FLUID SS-GALACTOSIDASE ASSAYS 55
2.5.3 WESTERN BLOT ANALYSES 55
2.5.3.1 SODIUM DODECYLSULFATE POLYACRYLAMIDE GEL ELECTROPHORESIS
(SDS-PAGE) _ 55
2.5.3.2 ELECTROPHERIC TRANSFER (SEMI DRY BLOT) AND IMMUNOLOGICAL
DETECTION OF PROTEINS 56
2.5.3.3 COOMASSIE STAINING OF POLYACRYLAMIDE GELS 56
2.5.3.4 SILVER STAINING OF POLYACRYLAMIDE GELS 56
2.5.3.5 PONCEAU-S STAINING 56
2.5.3.6 DRYING OF POLYACRYLAMIDE GELS 57
2.5.4 PROTEIN EXPRESSION 57
2.5.4.1 EXPRESSION OF RECOMBINANT PROTEIN IN E. COLI 57
2.5.4.2 TRANSFECTION AND EXPRESSION OF RECOMBINANT PROTEIN IN THE
BACULOVIRUS SYSTEM 57
2.5.5 ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) 59
2.5.6 MASS SPECTROMETRY (LIQUID CHROMATOGRAPHY ELECTROSPRAY IONISATION
TANDEM MS- LC ESI MS/MS) 59
2.5.6.1 SUPERNATANT EXTRACTION 59
RESULTS 61
3.1 IDENTIFICATION AND CLONING OF THE PUTATIVE S. RATTI HSP10 SEQUENCE
61
3.2 CHARACTERISATION OF THE & RATTI HSP10 TRANSCRIPT 62
3_5 IDENTIFICATION AND CLONING OF THE PUTATIVE S. ROTO HSP60 TRANSCRIPT
65
3.4 CHARACTERISATION OF S. RATTI HSP60 IL3 TRANSCRIPT 66
3.5 CHARACTERISATION OF PUTATIVE S. RATTI HSP10 AND HSP60 TRANSCRIPTS OF
PARASITIC FEMALES . 69
3.6 LOCALISATION OF PUTATIVE HSP10 AND HSP60 TRANSCRIPTS IN S. RATTI 13
BY IN SITU HYBRIDISATION 70
3.7 IDENTIFICATION OF HSP10 AND HSP60 PROTEINS IN SUPERNATANTS OF S.
RATTI 13 AND PARASITIC FEMALES 72
3.8 CHARACTERISATION OF THE PUTATIVE S. RATTI HSP10 AND HSP60 GENES 73
3.8.1 S. RATTI HSP10 AND HPS60 SOUTHERN BLOT ANALYSES 73
3.8.2 S. RATTI HSP10 AND HSP60 GENE ANALYSES 74
3.8.3 ANALYSES OF THE INTERGENIC REGION OF S. RATTI 75
IMAGE 4
TABLE OF CONTENTS
3.9 CHARACTERISATION OF THE S. RATTI HSP10 AND HSP60 AS PUTATIVE BINDING
PARTNERS USING TWO-HYBRID APPROACHES 78
3.9.1 MAMMALIAN TWO-HYBRID ANALYSIS 78
3.9.2 YEAST TWO-HYBRID ANALYSIS 81
3.10 DNA IMMUNISATION WITH RECOMBINANTLY EXPRESSED S. RATTI HSP10 85
3.11 PUTATIVE S. RATTI HSP60 EXPRESSION IN E. COLI 87
3.12 PUTATIVE S. RATTI HSP10 EXPRESSION IN THE BACULOVIRUS SYSTEM 88
4 DISCUSSION 89
4.1 IDENTIFICATION OF SRHSPIO AND SRHSP60 89
4.2 CHARACTERISATION OF SRHSPIO AND SRHSP60 90
43 CHARACTERISATION OF SRHSPIO AND SRHSP60 GENES 92
4.4 CHARACTERISATION OF SRHSPIO AND SRHSP60 AS BINDING PARTNERS _FEHLER!
TEXTMARKE NICHT DEFINIERT.
4.5 THE IMMUNOLOGICAL ROLE OF SRHSPIO 95
4.6 OUTLOOK 97
5 SUMMARY 98
6 APPENDIX 99
6.1 MAMMALIAN AND YEAST TWO-HYBRID S/-HSP60 BAIT-CONSTRUCTS 99
6. 1 . 1 SRHSPOO CDNA SEQUENCES OF THE MAMMALIAN TWO-HYBRID
BAIT-CONSTRUCTS 99
6.1.2 SRHSPLOCDNA SEQUENCES OFTHE MAMMALIAN TWO-HYBRID PREY-CONSTRUCTS
100
6.1.3 5/-HSP60 CDNA SEQUENCES OF THE YEAST TWO-HYBRID BAIT-CONSTRUCTS
101
6.1.4 SRHSPL 0 CDNA SEQUENCES OF THE YEAST TWO-HYBRID PREY-CONSTRUCTS
101
6.2 SRHSPIO AND SRHSP60 SOUTHERN BLOT PROBES 102
6.2.1 S/-HSP60 SOUTHERN BLOT PROBE CDNA SEQUENCES . 102
6.2.2 SRHSPIO SOUTHERN BLOT PROBE CDNA SEQUENCE 103
63 S/-HSP60 SUBFRAGMENT CDNA SEQUENCE FOR . COLI EXPRESSION EXCLUDING
THE
MITOCHONDRIA! LEADING SEQUENCE 103
6.4 S/-HSP60 CDNA AND AMINO ACID SEQUENCES INCLUDING THE 5 AND 3 UTR
104
6.5 SRHSPIO CDNA AND AMINO ACID SEQUENCES INCLUDING THE 5 AND 3 UTR
105
6.6 S/-HSP60 AND SRHSPIO PARASITIC FEMALE CDNA SEQUENCES 106
6.6.1 S/-HSP60 PARASITIC FEMALE CDNA SEQUENCE 106
6.6.2 SRHSPIO PARASITIC FEMALE CDNA SEQUENCE 108
6.7 SRHSP60 AND SRHSPIO GDNA SEQUENCES 109
IMAGE 5
TABLE OF CONTENTS
7 REFERENCES 110
8 ABBREVIATIONS AND UNITS 119
8.1 ABBREVIATIONS 119
8.2 UNITS 120
S3 AMINO ACIDS 121
|
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| author | Tazir, Yasmina |
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| ctrlnum | (OCoLC)560430219 (DE-599)DNB99845494X |
| dewey-full | 614.5934 |
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| dewey-ones | 614 - Forensic medicine; incidence of disease |
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| dewey-tens | 610 - Medicine and health |
| discipline | Medizin |
| edition | 1.Aufl. |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV026885334 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T23:20:47Z |
| institution | BVB |
| isbn | 9783835954717 3835954717 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-022411192 |
| oclc_num | 560430219 |
| open_access_boolean | |
| owner | DE-188 |
| owner_facet | DE-188 |
| physical | 123 S. Ill., graph. Darst. |
| publishDate | 2009 |
| publishDateSearch | 2009 |
| publishDateSort | 2009 |
| publisher | VVB Laufersweiler |
| record_format | marc |
| series2 | Edition scientifique |
| spelling | Tazir, Yasmina Verfasser (DE-588)139301178 aut Strongyloides ratti identification, isolation and characterisation of Heat Shock Protein 10 and Heat Shock Protein 60 eingereicht von Yasmina Tazir 1.Aufl. Giessen VVB Laufersweiler 2009 123 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Edition scientifique Zugl.: Giessen, Univ., Diss., 2009 antibodies cabt parasites cabt nematoda cabt strongyloididae cabt helminths cabt strongyloides ratti cabt strongyloides stercoralis cabt heat shock proteins cabt immune response cabt dna cloning cabt transcription cabt genome analysis cabt open reading frames cabt amino acid sequences cabt host parasite relationships cabt genes cabt immunity cabt (DE-588)4113937-9 Hochschulschrift gnd-content http://library.vetmed.fu-berlin.de/ResourceList/details/207755 lizenzfrei Abstract DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=022411192&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Tazir, Yasmina Strongyloides ratti identification, isolation and characterisation of Heat Shock Protein 10 and Heat Shock Protein 60 antibodies cabt parasites cabt nematoda cabt strongyloididae cabt helminths cabt strongyloides ratti cabt strongyloides stercoralis cabt heat shock proteins cabt immune response cabt dna cloning cabt transcription cabt genome analysis cabt open reading frames cabt amino acid sequences cabt host parasite relationships cabt genes cabt immunity cabt |
| subject_GND | (DE-588)4113937-9 |
| title | Strongyloides ratti identification, isolation and characterisation of Heat Shock Protein 10 and Heat Shock Protein 60 |
| title_auth | Strongyloides ratti identification, isolation and characterisation of Heat Shock Protein 10 and Heat Shock Protein 60 |
| title_exact_search | Strongyloides ratti identification, isolation and characterisation of Heat Shock Protein 10 and Heat Shock Protein 60 |
| title_full | Strongyloides ratti identification, isolation and characterisation of Heat Shock Protein 10 and Heat Shock Protein 60 eingereicht von Yasmina Tazir |
| title_fullStr | Strongyloides ratti identification, isolation and characterisation of Heat Shock Protein 10 and Heat Shock Protein 60 eingereicht von Yasmina Tazir |
| title_full_unstemmed | Strongyloides ratti identification, isolation and characterisation of Heat Shock Protein 10 and Heat Shock Protein 60 eingereicht von Yasmina Tazir |
| title_short | Strongyloides ratti |
| title_sort | strongyloides ratti identification isolation and characterisation of heat shock protein 10 and heat shock protein 60 |
| title_sub | identification, isolation and characterisation of Heat Shock Protein 10 and Heat Shock Protein 60 |
| topic | antibodies cabt parasites cabt nematoda cabt strongyloididae cabt helminths cabt strongyloides ratti cabt strongyloides stercoralis cabt heat shock proteins cabt immune response cabt dna cloning cabt transcription cabt genome analysis cabt open reading frames cabt amino acid sequences cabt host parasite relationships cabt genes cabt immunity cabt |
| topic_facet | antibodies parasites nematoda strongyloididae helminths strongyloides ratti strongyloides stercoralis heat shock proteins immune response dna cloning transcription genome analysis open reading frames amino acid sequences host parasite relationships genes immunity Hochschulschrift |
| url | http://library.vetmed.fu-berlin.de/ResourceList/details/207755 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=022411192&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT taziryasmina strongyloidesrattiidentificationisolationandcharacterisationofheatshockprotein10andheatshockprotein60 |