Molecular interactions of the malaria parasite Plasmodium falciparum during the sexual reproduction in the mosquito midgut:
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| Main Author: | |
|---|---|
| Format: | Thesis/Dissertation Book |
| Language: | English |
| Published: |
2012
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| Subjects: | |
| Links: | https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-72403 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025245714&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| Item Description: | Zsfassung in dt. und engl. Sprache |
| Physical Description: | VI, 145 Bl. Ill., graph. Darst. |
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| 100 | 1 | |a Simon, Nina Monica |e Verfasser |0 (DE-588)1025516826 |4 aut | |
| 245 | 1 | 0 | |a Molecular interactions of the malaria parasite Plasmodium falciparum during the sexual reproduction in the mosquito midgut |c von Nina Monica Simon |
| 246 | 1 | 3 | |a Molekulare Wechselwirkungen des Malariaparasiten Plasmodium falciparum während der sexuellen Fortpflanzung im Mitteldarm der Mücke |
| 264 | 1 | |c 2012 | |
| 300 | |a VI, 145 Bl. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
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| 500 | |a Zsfassung in dt. und engl. Sprache | ||
| 502 | |a Würzburg, Univ., Diss., 2012 | ||
| 650 | 0 | 7 | |a Sexuelle Entwicklung |0 (DE-588)4121668-4 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Malaria |0 (DE-588)4037197-9 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Plasmodium falciparum |0 (DE-588)4250521-5 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Plasmodium falciparum |0 (DE-588)4250521-5 |D s |
| 689 | 0 | 1 | |a Sexuelle Entwicklung |0 (DE-588)4121668-4 |D s |
| 689 | 0 | 2 | |a Malaria |0 (DE-588)4037197-9 |D s |
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Record in the Search Index
| DE-BY-TUM_katkey | 1871901 |
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| adam_text | IMAGE 1
TABLE O F CONTENTS
TABLE OF CONTENTS
1 INTRODUCTION 1
1.1 MALARIA 1
1.2 THE MALARIA PATHOGEN PLASMODIUM FALCIPARUM 3
1.2.1 LIFE CYCLE 3
1.2.2 PLASMODIUM FALCIPARUM SEXUAL STAGES 5
1.3 TRANSMISSION BLOCKING VACCINE 7
1.4 SEXUAL STAGE-SPECIFIC PROTEINS OF THE MALARIA PARASITE 9
1.5 PFCCP MULTI-ADHESION DOMAIN PROTEINS 12
1.6 THE MALARIA PARASITE AND THE HUMAN COMPLEMENT IN THE MOSQUITO MIDGUT
15
1.6.1 SEXUAL REPRODUCTION O F MALARIA PARASITES IN THE MOSQUITO MIDGUT
15
1.6.2 THE HUMAN COMPLEMENT SYSTEM 16
1.6.3 REGULATION O F THE ALTERNATIVE PATHWAY O F COMPLEMENT 17
1.6.4 PROPERTIES O F COMPLEMENT FACTOR C3 AND FACTOR H 19
1.6.5 PATHOGENS BIND FACTOR H FOR COMPLEMENT INACTIVATION 2 1
1.7 AIM O F THE STUDY 22
2 MATERIALS AND METHODS 24
2.1 MATERIALS 24
2.1.1 INSTRUMENTS 24
2.1.2 CHEMICALS AND CONSUMABLES 25
2.1.3 KITS 26
2.1.4 BUFFERS AND SOLUTIONS 26
2.1.5 MEDIUM AND AGAR PLATES 28
2.1.6 PARASITE AND BACTERIAL CELL LINES 29
2.1.7 PLASMIDS 30
2.1.8 PROTEIN LADDERS 30
2.1.9 ANTIBODIES AND PROTEINS 32
2.1.10 PROTEIN IDENTIFICATION NUMBERS (PLASMODB AND UNIPROT) 34
2.1.11 BIOINFORMATIC SOURCES AND COMPUTER PROGRAMS 35
2.2 METHODS 35
2.2.1 CELL BIOLOGY 35
2.2.1.1 CULTIVATION AND STORAGE O F BACTERIA 35
2.2.1.2 TRANSFORMATION O F COMPETENT BACTERIAL CELLS 35
2.2.1.3 CULTURE OF PLASMODIUM FALCIPARUM 36
IV
HTTP://D-NB.INFO/1026943868
IMAGE 2
TABLE O F CONTENTS
2.2.1.4 DETERMINATION OF PARASITEMIA AND GAMETOCYTEMIA 37
2.2.1.5 INDUCTION OF GAMETOGENESIS 39
2.2.1.6 GAMETOGENESIS INHIBITION ASSAY (GIA) 40
2.2.1.7 GAMETOCYTE PURIFICATION 40
2.2.1.8 INDIRECT IMMUNOFLUORESCENCE ASSAY (IFA) 4 1
2.2.2 PROTEIN BIOCHEMICAL METHODS 42
2.2.2.1 EXPRESSION O F RECOMBINANT PROTEINS 42
2.2.2.2 PURIFICATION O F RECOMBINANT PROTEINS 42
2.2.2.3 SDS POLYACRYLAMIDE GEL ELECTROPHORESIS (PAGE) 43
2.2.2.4 SDS GEL STAINING, MEASUREMENT O F THE PROTEIN CONCENTRATION AND
CONSERVATION 44
2.2.2.5 WESTERN BLOT ANALYSIS 45
2.2.2.6 INTERACTION STUDIES 46
2.2.2.7 IMMUNIZATION OF MICE AND GENERATION OF ANTIBODIES 47
2.2.2.8 PREPARATION AND HANDLING OF HUMAN SERUM 48
2.2.2.9 DETERMINATION O F COMPLEMENT ACTIVITY 48
2.2.2.10 CO-FACTOR ACTIVITY ASSAY 49
2.2.2.11 CELL TREATMENT FOR INVESTIGATION O F CELL BINDING COMPLEMENT
FACTORS 49
2.2.3 MOSQUITO REARING AND FEEDING TECHNIQUES 49
2.2.3.1 REARING OF ANOPHELES STEPHENSI MOSQUITOES 50
2.2.3.2 EX VIVO FEEDING O F MOSQUITOES 50
2.2.3.3 DISSECTION O F MOSQUITO MIDGUTS 5 1
3 RESULTS 52
3.1 SEXUAL STAGE PROTEINS O F THE MALARIA PARASITE 52
3.1.1 ORTHOLOGS AND ADHESION DOMAINS O F PFCCP PROTEINS 52
3.1.2 MODULES O F THE PFCCP PROTEINS 54
3.1.3 MOLECULAR INTERACTIONS BETWEEN SEXUAL STAGE PROTEINS O F
PLASMODIUM FALCIPARUM 54
3.1.3.1 CO-IMMUNOPRECIPITATION STUDIES IN GAMETOCYTES AND ACTIVATED
GAMETOCYTES 55
3.1.3.2 CO-ELUTION BINDING ASSAYS WITH RECOMBINANT PFCCP PROTEINS 59
3.1.3.3 INTERACTION STUDIES OF ENDOGENOUS PROTEINS WITH RECOMBINANT 62
PROTEINS
3.1.3.4 IDENTIFICATION O F AN ADDITIONAL INTERACTION PARTNER O F THE MPC
63
3.1.4 PROCESSING OF PFCCP PROTEINS 65
V
IMAGE 3
TABLE O F CONTENTS
3.2 HUMAN COMPLEMENT IN THE MOSQUITO MIDGUT 67
3.2.1 ACTIVITY O F THE COMPLEMENT SYSTEM IN THE MOSQUITO MIDGUT 67
3.2.2 EXAMINATION O F CLASSICAL PATHWAY ACTIVATION 68
3.2.3 BINDING O F HUMAN COMPLEMENT FACTOR C3 TO MALARIA PARASITES 70
3.2.4 IMPACT O F THE ACTIVE HUMAN COMPLEMENT ON PARASITE GAMETOGENESIS
72
3.2.5 HUMAN FACTOR H IN THE MOSQUITO MIDGUT 76
3.2.6 BINDING STUDIES OF FH AND FH FAMILY PROTEINS TO MALARIA PARASITES
77
3.2.6.1 INDIRECT IMMUNOFLUORESCENCE ASSAY O F FH BINDING 77
3.2.6.2 SERUM ABSORBANCE ASSAY WITH MALARIA PARASITES 79
3.2.6.3 BINDING STUDIES OF FH FAMILY PROTEINS TO MALARIA PARASITES BY
WESTERN BLOTTING 80
3.2.6.4 PEPTIDE BINDING ASSAY O F RECOMBINANT FH-PROTEINS ON ACTIVATED
GAMETOCYTES 82
3.2.7 COFACTOR ACTIVITY O F FH ON THE SURFACE O F AGC 84
3.2.8 INFLUENCE O F HEPARIN ON C3B AND FH BINDING 84
3.3 INTERACTION PARTNER O F FH ON THE SURFACE O F ACTIVATED GAMETOCYTES
86
3.3.1 IDENTIFICATION O F THE FH RECEPTOR PROTEIN 86
3.3.2 EXPRESSION AND LOCALIZATION STUDIES O F PFGAP50 IN SEXUAL STAGE
PARASITES.... 88
3.3.3 INTERACTION STUDIES WITH RECOMBINANT PFGAP50 PROTEINS AND FH
PEPTIDES.. 90
3.4 IFA CONTROLS 9 1
4 DISCUSSION 92
4.1 SEXUAL STAGE SURFACE PROTEINS O F THE MALARIA PARASITE 92
4.2 THE HUMAN COMPLEMENT SYSTEM AND MALARIA PARASITES IN THE MOSQUITO
MIDGUT 101
5 FUTURE PERSPECTIVES I L L
6 SUMMARY 113
7 ZUSAMMENFASSUNG 115
8 REFERENCES 117
9 APPENDIX 138
9.1 ABBREVIATIONS 138
9.2 LIST O F FIGURES 140
9.3 LIST O F TABLES 142
9.4 CURRICULUM VITAE 143
9.5 PUBLICATIONS 145
VI
|
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| language | English |
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| physical | VI, 145 Bl. Ill., graph. Darst. |
| psigel | ebook |
| publishDate | 2012 |
| publishDateSearch | 2012 |
| publishDateSort | 2012 |
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| spellingShingle | Simon, Nina Monica Molecular interactions of the malaria parasite Plasmodium falciparum during the sexual reproduction in the mosquito midgut Sexuelle Entwicklung (DE-588)4121668-4 gnd Malaria (DE-588)4037197-9 gnd Plasmodium falciparum (DE-588)4250521-5 gnd |
| subject_GND | (DE-588)4121668-4 (DE-588)4037197-9 (DE-588)4250521-5 (DE-588)4113937-9 |
| title | Molecular interactions of the malaria parasite Plasmodium falciparum during the sexual reproduction in the mosquito midgut |
| title_alt | Molekulare Wechselwirkungen des Malariaparasiten Plasmodium falciparum während der sexuellen Fortpflanzung im Mitteldarm der Mücke |
| title_auth | Molecular interactions of the malaria parasite Plasmodium falciparum during the sexual reproduction in the mosquito midgut |
| title_exact_search | Molecular interactions of the malaria parasite Plasmodium falciparum during the sexual reproduction in the mosquito midgut |
| title_full | Molecular interactions of the malaria parasite Plasmodium falciparum during the sexual reproduction in the mosquito midgut von Nina Monica Simon |
| title_fullStr | Molecular interactions of the malaria parasite Plasmodium falciparum during the sexual reproduction in the mosquito midgut von Nina Monica Simon |
| title_full_unstemmed | Molecular interactions of the malaria parasite Plasmodium falciparum during the sexual reproduction in the mosquito midgut von Nina Monica Simon |
| title_short | Molecular interactions of the malaria parasite Plasmodium falciparum during the sexual reproduction in the mosquito midgut |
| title_sort | molecular interactions of the malaria parasite plasmodium falciparum during the sexual reproduction in the mosquito midgut |
| topic | Sexuelle Entwicklung (DE-588)4121668-4 gnd Malaria (DE-588)4037197-9 gnd Plasmodium falciparum (DE-588)4250521-5 gnd |
| topic_facet | Sexuelle Entwicklung Malaria Plasmodium falciparum Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-72403 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025245714&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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