New Perspectives in Science Education

Edition 14

Accepted Abstracts

The Yeast Two Hybrid Screening as a Teaching Tool

Esra Seyran, Department of Molecular Biology and Genetics, Cumhuriyet University (Turkey)

Abstract

The laboratory courses based on research has proved to attract and encourage student interests. Here we developed a semester long laboratory course to accompany the theoretical courses of biochemical genetics or microbial genetics. The research project was designed to make students familiar with basic concepts and techniques of microbiology, recombinant DNA, transcription, translation, bioinformatics, and proteomics. The investigation of protein-protein interactions has cross disciplinary importance. The Yeast Two Hybrid (Y2H) technique is one of the established tools used for the investigation of protein-protein interactions.  In general, in any Yeast Two Hybrid experiment, a protein of interest is fused to a DNA-binding domain, and then this protein is used to screen a library of cDNA clones that are fused to an activation domain. The yeast two hybrid is a system designed to identify genes in a cDNA library encoding proteins that are physically associated with the protein of interest in vivo. We have used the Yeast Two Hybrid system to understand the protein-protein interactions in riceblast pathosystem. Plant disease control is an area where the field of protein-protein interactions has the potential to make a significant and immediate impact through the identification of proteins involved in disease resistance. Genetic analyses of plant-pathogen interactions have revealed that resistance reactions are mainly controlled by two dominant loci; avirulence gene in the pathogen and a corresponding resistance gene in the plant.  The absence of either member of the gene pair results in the development of disease symptoms. In order to fully exploit the potential of yeast two hybrid system, we constructed a rice cDNA library and screened this rice cDNA library across the avr-pita protein of the rice pathogen. The protein sequence of positive interactors were deposited at National Center for Biotechnology Information (NCBI) database. The main objective of the course was to make students experience the excitement of scientific discovery.

Keywords: Molecular biology, yeast two hybrid, research based teaching;

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