Molecular genetic investigation of the variability of the GTPase activating protein- (GAP-) related domain of the tuberous sclerosis-2 (TSC2) gene in TSC patients and healthy subjects [Elektronische Ressource] / vorgelegt von Karin Louise Zügge geb. Gierke

Zentrum für Kinderheilkunde und Jugendmedizin Albert-Ludwigs-Universität Freiburg, Germany MOLECULAR GENETIC INVESTIGATION OF THE VARIABILITY OF THE GTPase ACTIVATING PROTEIN- (GAP-) RELATED DOMAIN OF THE TUBEROUS SCLEROSIS - 2 (TSC2) GENE IN TSC PATIENTS AND HEALTHY SUBJECTS Inaugural - Dissertation zur Erlangung des Medizinischen Doktorgrades der Medizinischen Fakultät der Albert-Ludwigs-Universität Freiburg i. Br. Vorgelegt 2002 von Karin Louise Zügge geb. Gierke geboren in Madison, Wisconsin/U.S.A. Dekan: Prof. Dr. med. J. Zentner 1. Gutachter: Prof. Dr. med. L.B. Zimmerhackl 2. Gutachter: Prof. Dr. med. E. Schulz Jahr der Promotion: 2004 i Table of Contents 1 Introduction 1-12 1.1 History 1 1.2 Clinical characteristics 3 1.3 Pathogenesis 5 1.4 Genetics 6 1.5 Protein products 7 1.6 The clinical variability predicament 10 1.7 The Freiburg-Heidelberg project 11 1.8 Objectives and research questions 12 2 Materials and Methods 13-22 2.1 Materials 13-15 2.1.1 Apparatus and equipment 13 2.1.2 Buffers and solutions 13 2.1.3 Chemicals, enzymes, kits and size markers 14 2.1.4 Databases and software 14 2.1.5 Media and cell cultures 14 2.1.6 Oligonucleotides and DNA-sequencing 15 2.2 Methods 15-22 2.2.1 Probands 15 2.2.1.1 TSC patients and families 15 2.2.1.2 Healthy population 15 2.2.2 Cell culture and transformation from peripheral blood samples 16 2.2.