Membrane proteins in the outer membrane of plastids and mitochondria [Elektronische Ressource] / vorgelegt von Iryna Ilkavets

Membrane proteins in the outer membrane of plastids and mitochondria vorgelegt von Iryna Ilkavets Dissertation der Fakultät für Biologie der Ludwig-Maximilians-Universität München München 19.12.2005 Gutachter: 1. Prof. Dr. J. Soll 2. PD Dr. J. Meurer Date of the exam: 13.02.2006 1Contents Sumary 4 Zusamenfasung 5 Abbreviations 6 1. Introduction 7 2. Materials and methods 9 2.1 Bacterial strains 9 2.2 Plant material 2.3 DNA methods 9 2.3.1 Isolation of genomic DNA from Arabidopsis thaliana 10 2.3.2 Polymerase Chain Reaction 10 2.3.3 Southern hybridisation 10 2.4 Cloning 11 2.4.1 Conventional cloning 11 2.4.2 Site directed mutagenesis 2.4.3 GATEWAY 2.5 RNA methods 12 2.5.1 isolation from plant material 12 2.5.2 cDNA synthesis 2.5.3 Semi-quantitative RT-PCR 12 2.5.4 cDNA macroarray analysis of wild type and Atoep16.1-p knockout mutant 12 2.5.5 Affymetrix genechip analysis 13 2.6 Overexpression of recombinant proteins and antibody purification 13 2.6.1 Heterologous expression of proteins in E.coli 14 2.6.2 Inclusion bodies preparation 15 2.6.3 Purification of overexpressed protein 15 2.6.4 Antibody production 15 2.7 GFP, RFP-fusion protein analysis 16 2.7.