Asian Journal of Microbiology, Biotechnology & Environmental Sciences Paper

Vol.12, Issue 3, 2010; Page No.(639-645)

MOLECULAR CHARACTERIZATION OF YEAST AND THE MUTANTS DEVELOPED BY ETHIDIUM BROMIDE MUTATION

HARINI KUMAR, ANNAPURNA S. AGASTHYA, K. RIJESH, AMRUTHA MURUGESH, SANDEEP BOSE AND NITHYA PRIYA

Abstract

Saccharomyces cerevisiae gained particular attention in the 1990s as it has served as an important scientific model and tool, and was the first eukaryotic organism to have its genome sequenced. Mutations may be induced by exposure to ultraviolet rays and alpha, beta, gamma, and X radiation, by extreme changes in temperature, and by certain mutagenic chemicals such as nitrous acid, nitrogen mustard, and chemical substitutes for portions of the nucleotide subunits of genes. Ethidium bromide (EtBr), is an intercalating agent commonly used as a fluorescent tag (nucleic acid stain) in molecular biology laboratories for techniques such as agarose gel electrophoresis. Ethidium bromide may be a strong mutagen. The molecular markers are one of the versatile tools in Molecular Biology and Genetic Engineering. Molecular genetic techniques can be used to discriminate between yeast strains that have similar physiological characteristics. In the present study, induced mutation was carried out with interchelating agent like ethidium bromide with four different concentrations, 0.25, 0.50, 0.75 and 1 μg/ml. Random Amplified Polymorphic DNA (RAPD) analysis was employed to characterize yeast isolates. Five Saccharomyces strains were subjected to RAPD analysis using nine primers, selected from the kits of Chromous Biotech. Dendrogram was constructed according to Wards method based on the scorable bands using Statistica software. Cluster analysis of dendrograms revealed that all the five yeast isolates formed two major clusters.

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