Microbial Gene Technology

Abstract

Introduction to selected prokaryotic and eukaryotic model microorganisms and application of methods which are used as tools for answering questions in molecular biology.

Objective

Knowledge about the introduced microorganisms, about the methods which are introduced in the course and their application to answer general questions in molecular biology.

Content

- Isolation of temperature-sensitive mutations in an essential PNG gene of the yeast Saccharomyces cerevisiae by low fidelity PCR and in vivo recombination. -Characterization of the obtained mutations by DNA sequencing. -Complementation of mutations in non-essential PNG genes by the homologous human genes and biochemical analysis of the N-glycans of a reporter protein in various PNG mutants. -Test of various PNG genes for genetic interaction according to a protocol of a high-throughput screen for synthetic lethal mutations in S. cerevisiae. - PCR amplification and cloning of genomic DNA of the soybean symbiont Bradyrhizobium japonicum - Site-directed mutagenesis (“Quikchange" method) - Overexpression of regulatory genes and affinity purification of proteins - Protein-DNA interaction studies

Resources