S. cerevisiae yeasts were obtained from alcohol factory’s effluent
A total of 52 microorganism samples from alcohol factory’s effluent after preparation and cultivation of these samples on the complete medium for yeast growth, were isolated and their total DNA were extracted. Then the PCR method was performed to examine Zrt1 and Fet4 genes in these samples. Four samples expressing Zrt1 and Fet4 genes which illustrated fragment patterns of 2102 bp and 165 bp on 1.5% agarose gel in relation with Zrt1 and Fet4 genes, respectively, were identified (figure 1A). Subsequently, after isolation of samples which express Zrt1 and Fet4 genes, the yeast strains of these four samples were identified using PCR of ITS1 and ITS4 primers. Then PCR products were electrophoresed and results observed on 1.5% agarose gel and three out of four samples illustrated fragment patterns in relation with ITS, representing these three samples as yeast strains (figure1B). Then related fragments were verified by sequencing and the results identified two samples as S. cerevisiae yeasts (figure 1C). All results of gel electrophoresis as well as phylogenetic tree of sequenced samples were illustrated in figure 1 (to observe the results of sample sequencing see the supplementary pdf file 1).