No abstract

Low frequency, integrative transformation of three fertile hermaphroditic strains of Magnaporthe grisea has been achieved using plasmid pAN7-1 and cosmid pAN7-2, which contain an Escherichia coli hygromycin B phosphotransferase gene linked to Aspergillus nidulans regulatory sequences.

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Four different types of endo-fl-l,4-glucanase active bottom-fermenting brewer's yeast strains were constructed using recombinant DNA technology. To study the effects of different promoters, copy numbers and integration sites, the eoll gene of the filamentous fungus Trichoderma reesei was inserted between the promoter and terminator regions of either the PGK1 or ADH1 gene of yeast. The eoll gene was transferred to the industrial brewer's yeast on a multicopy plasmid or alternatively integrated into the LEU2, PGK1 or A D H 1 locus of the yeast. Integration into the PGK1 or A D H 1 locus did not affect the brewing properties of the yeast or the quality of the finished beer. Integration into the LEU2 locus, however, decreased the metabolic activity of yeast and prolonged fermentation was needed. In pilot brewing conditions the PGK1 promoter was stronger than that of ADH1. Even a single copy of the eoH gene in the PGK1 integrant strains gave rise to sufficient enzyme activity for the hydrolysis even of unusually high total amounts of fl-glucans in worts.