Molekulare Pathogenese von CADASIL: Mechanismen der Notch3 Rezeptoraggregation
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | German |
| Veröffentlicht: |
2009
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| Schlagworte: | |
| Online-Zugang: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:19-97597 Inhaltsverzeichnis |
| Beschreibung: | 56 Bl. Ill., graph. Darst. |
Internformat
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| 245 | 1 | 0 | |a Molekulare Pathogenese von CADASIL |b Mechanismen der Notch3 Rezeptoraggregation |c von Anna Magdalena Karpinska |
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Datensatz im Suchindex
| _version_ | 1804138844724920321 |
|---|---|
| adam_text | Titel: Molekulare Pathogenese von CADASIL
Autor: Karpinska, Anna
Jahr: 2009
Inhaltsverzeichnis
Inhaltsverzeichnis
ABKÜRZUNGSVERZEICHNIS.........................................................................................1
1 EINLEITUNG..................................................................................................................3
1.1 Klinischer Überblick und molekulare Grundlagen der Erkrankung.................3
1.2 Fragestellung der Arbeit......................................................................................6
2 MATERIAL UND METHODIK........................................................................................8
2.1 Material..................................................................................................................8
2.1.1 Allgemeine Ausrüstung und Geräte...............................................................8
2.1.2 Reagenzien....................................................................................................9
2.1.3 Puffer, Lösungen..........................................................................................12
2.1.3.1 DNA..........................................................................................................12
2.1.3.2 Zellkultur....................................................................................................15
2.1.3.3 Proteine.....................................................................................................16
2.2 Methodik..............................................................................................................17
2.2.1 DNA-Methoden............................................................................................17
2.2.1.1 Transformation von Bakterien...................................................................17
2.2.1.2 Stammkonservierung................................................................................17
2.2.1.3 Maxi-Preparation.......................................................................................17
2.2.1.4 Konzentrationsbestimmung.......................................................................18
2.2.1.5 Kontrolle der resynthetisierten Plasmide per Restriktionsverdau..............19
2.2.1.6 Sequenz-PCR...........................................................................................19
2.2.2 Zellkultur-Methoden.....................................................................................20
2.2.2.1 Kultivierung der Zelllinie............................................................................20
2.2.2.2 Transfekuon..............................................................................................21
2.2.2.3 Zelllyse......................................................................................................21
2.2.3 Protein-Methoden.........................................................................................21
2.2.3.1 Antikörper..................................................................................................21
2.2.3.2 Proteinbestimmung...................................................................................22
2.2.3.3 SDS-Page.................................................................................................22
2.2.3.4 Westem Blot..............................................................................................23
2.2.3.5 Co-Immunopräzipitation (IP).....................................................................23
2.2.3.6 Zeitreihen..................................................................................................24
2.2.3.7 SIFT scanning for intensely fluorescent targets........................................24
Inhaltsverzeichnis
3 ERGEBNISSE..............................................................................................................27
3.1 Notch3-Konstrukte.............................................................................................27
3.2 Expressionsnachweis von Notch3 in HEK293 Zellen......................................28
3.3 Immunopräzipitation zur Untersuchung einer homophilen N3EC Interaktion 29
3.4 Immunopräzipitation zur Untersuchung einer homophilen N3FL Interaktion 32
3.5 Untersuchung eines Ca2* Einflusses auf die N3EC- Interaktion......................35
3.6 Zeitabhängige Bildung höhermolekularer ß-ME resistenter N3-Komplexe... 36
3.7 Untersuchung der N3EC4nteraktion mittels SIFT.............................................39
4 DISKUSSION...............................................................................................................42
4.1 Zusammenfassung der Ergebnisse..................................................................42
4.1.1 Co-Immunopräzipitationen...........................................................................42
4.1.2 Zeitabhängigkeit der Komplexbildung..........................................................43
4.1.3 SIFT - scanning for intensely fluorescent targets.........................................43
4.2 Evaluierung der Versuchsbedingungen...........................................................44
4.2.1 Fehlende Proteinaufreinigung......................................................................44
4.2.2 Artifizielle Überexpression............................................................................45
4.2.3 Vorteile des SIFT gegenüber anderen Proteinnachweismethoden..............45
4.2.4 Indirekte Fluoreszenzmarkierung im SIFT....................................................45
4.3 Relevanz der Ergebnisse hinsichtlich der CADASIL-Pathogenese...............46
4.3.1 Strukturänderungen des N3-Rezeptors........................................................46
4.3.2 Störungen des Notch3-Signalweges............................................................47
4.3.3 Neomorpher Effekt der CADASIL-Mutationen..............................................48
4.3.4 Bedeutung der zeitabhängigen Komplexbildung für die Pathogenese.........48
5 ZUSAMMENFASSUNG................................................................................................50
6 ABBILDUNGSVERZEICHNIS......................................................................................52
7 LITERATURVERZEICHNIS.........................................................................................53
8 LEBENSLAUF..............................................................................................................56
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| dewey-ones | 616 - Diseases |
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| dewey-tens | 610 - Medicine and health |
| discipline | Medizin |
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| indexdate | 2024-07-09T21:34:34Z |
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| language | German |
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| spelling | Karpinska, Anna 1982- Verfasser (DE-588)137808127 aut Molekulare Pathogenese von CADASIL Mechanismen der Notch3 Rezeptoraggregation von Anna Magdalena Karpinska 2009 56 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Univ., Diss., 2009 (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:19-97597 http://edoc.ub.uni-muenchen.de/9759/ Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:19-97597 Resolving-System HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=017329155&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Karpinska, Anna 1982- Molekulare Pathogenese von CADASIL Mechanismen der Notch3 Rezeptoraggregation |
| subject_GND | (DE-588)4113937-9 |
| title | Molekulare Pathogenese von CADASIL Mechanismen der Notch3 Rezeptoraggregation |
| title_auth | Molekulare Pathogenese von CADASIL Mechanismen der Notch3 Rezeptoraggregation |
| title_exact_search | Molekulare Pathogenese von CADASIL Mechanismen der Notch3 Rezeptoraggregation |
| title_full | Molekulare Pathogenese von CADASIL Mechanismen der Notch3 Rezeptoraggregation von Anna Magdalena Karpinska |
| title_fullStr | Molekulare Pathogenese von CADASIL Mechanismen der Notch3 Rezeptoraggregation von Anna Magdalena Karpinska |
| title_full_unstemmed | Molekulare Pathogenese von CADASIL Mechanismen der Notch3 Rezeptoraggregation von Anna Magdalena Karpinska |
| title_short | Molekulare Pathogenese von CADASIL |
| title_sort | molekulare pathogenese von cadasil mechanismen der notch3 rezeptoraggregation |
| title_sub | Mechanismen der Notch3 Rezeptoraggregation |
| topic_facet | Hochschulschrift |
| url | http://edoc.ub.uni-muenchen.de/9759/ https://nbn-resolving.org/urn:nbn:de:bvb:19-97597 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=017329155&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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