RESUMO
Replacement of the regulatory and secretory signals of the alpha-amylase gene (AMY) from Bacillus amylolique-faciens with the complete yeast pheromone alpha-factor prepro region (MF alpha 1p) resulted in increased levels of extracellular alpha-amylase production in Saccharomyces cerevisiae. However, the removal of the (Glu-Ala)2 peptide from the MF alpha 1 spacer region (Lys-Arg-Glu-Ala-Glu-Ala) yielded decreased levels of extracellular alpha-amylase.
Assuntos
Bacillus/enzimologia , Peptídeos/genética , Saccharomyces cerevisiae/enzimologia , alfa-Amilases/metabolismo , Bacillus/genética , Sequência de Bases , Regulação Enzimológica da Expressão Gênica , Fator de Acasalamento , Dados de Sequência Molecular , Saccharomyces cerevisiae/genética , Transcrição Gênica , Transfecção , alfa-Amilases/genéticaRESUMO
K2 neutral strain Saccharomyces cerevisiae USM12 was identified and characterized. This strain carried an M double-stranded RNA (dsRNA) genome encoding for resistance to K2 toxin. The M dsRNA was larger than the K2 killer yeast M dsRNA and homoduplex analysis of denatured and reannealed K2 neurtal M dsRNA revealed an inverted duplication. Heteroduplex analysis showed that two thirds of the K2 M genome had homology with the K2 neutral M genome. Hybridization showed that the USM12 M dsRNA had significant homology with the K2 M dsRNA. Protein profiles of extracellular proteins from USM12 and a cured strain indicated that USM12 did not secrete any toxin. This is the first time that a K2 neutral yeast strain has been characterized.
