Both an α-amylase gene of Bacillus subtilis RSKK246 and a gene encoding β−glucanase from B. subtilis RSKK243 were cloned and expressed in both E. coli XL1-Blue MRF and B. subtilis YB886 by using the vectors pUC18 and pUB110 respectively. | Turk J Agric For 25 (2001) 45-49 © TÜB‹TAK Heterologous Expression of Genes in the Yeast Saccharomyces cerevisiae Numan ÖZCAN Çukurova University, Agricultural Faculty, Department of Animal Science, Adana - TURKEY Received: Abstract : Both an α-amylase gene of Bacillus subtilis RSKK246 and a gene encoding β−glucanase from B. subtilis RSKK243 were cloned and expressed in both E. coli XL1-Blue MRF and B. subtilis YB886 by using the vectors pUC18 and pUB110 respectively. These genes were also cloned into the E. coli-yeast shuttle vectors pRS406 and pRS416 for transfer into the yeast Saccharomyces cerevisiae. These constructs carrying α-amylase and β-glucanase, genes which were cloned into the pRS406 vector, were expressed by integration into the yeast chromosome. These genes were also expressed in the yeast by autonomous replication of the ARS/CEN plasmid pRS416. This work demonstrates that genes belonging to B. subtilis can replicate in both E. coli and in the yeast S. cerevisiae. Key Words : α-amylase, β−glucanase, Saccharomyces cerevisiae, Bacillus subtilis, gene cloning Maya Saccharomyces cerevisiae’yede Farkl› Genlerin Ekspresyonu Özet : Bacillus subtilis RSKK246 ya ait α-amilaz ve B. subtilis RSKK243’e ait β−glukanaz enzimlerini kodlayan genler klonlanarak E. coli XL1-Blue MRF ve B. subtilis YB886’da s›ras›yla pUC18 ve pUB110 vektörleri kullan›larak ekspres edildi. Bu genler E. colimaya shuttle vektörleri olan pRS406 ve pRS416’ya klonlanarak maya Saccharomyces cerevisiae’ye de transfer edildi. a-amilaz ve β− glukanaz genlerini pRS406 vektöründe tafl›yan bu yap›lar maya kromozomuna entegre edilerek ekspres edildi. Bu genler ayr›ca ARS/CEN bölgesi tafl›yan pRS416 replikasyon plasmidi ile mayada replike edilerek ekspres edildi. Bu çal›flma B. subtilis’e ait genlerin E. coli ve maya S. cerevisiae’ye de enzim aktivitesini kaybetmeden ekspres olunabilece¤ini ispatlam›flt›r. Anahtar Sözcükler : α-amilaz, β−glukanaz, Saccharomyces cerevisiae, Bacillus .
