Construction of recombinant yeast(Saccharomyces cerevisiae) producing ß(1,3) glucanase as a fish feed addictives

In this study, ß(1,3) glucanase gene was digested by Sacl restriction enzyme and obtained by purification from pETG11(pUC18+ß(1,3) glucanase recombinant vector. It was then transfered to the pRS416 vector which was digested by Sacl restriction enzyme and recombinant vector was constructed. The new vector is called pRS416G, and a replicative vector was also developed for the yeast (Saccharomyces cerevisiae). After that pRS416G recombinant vector was transferred to Saccharomyces cerevisiae using electroporation. The PCR reaction, used as a template by DNA of recombinant Saccharomyces cerevisiae/pRS416G, was studied in agarose gel of 0.8%, and a DNA band of ß(1,3) glucanase gen, approximately 1.9 kbp in size, was observed. In addition, the yellow zone was observed in recombinant yeast as a result of zymogram analysis, suggesting that ß(1,3) gene functions.