Author's Organization：National Food Research Institute, Ministry of Agriculture, Forestry and Fisheries Institute of Food Research and Product Development, Kasetsart University Department of Agricultural Chemistry, The University of Tokyo:(Present address) Department of Applied Biological Science, Tokyo University of Agriculture and Technology Department of Agricultural Chemistry, The University of Tokyo Department of Agricultural Chemistry, The University of Tokyo:(Present address) Nagaoka University of Technology National Food Research Institute, Ministry of Agriculture, Forestry and Fisheries Department of Agricultural Chemistry, The University of Tokyo
We have developed a genetic transformation system for a freeze-tolerant yeast Kluyveromyces thermotolerans. K. thermotolerans spheroplasts could be transformed with a YPp-type vector containing an autonomously replicating (ARS) of Saccharomyces cerevisiae. However, transformation with a Yep-type vector containing a replication origin of S. cerevisiae 2μM DNA was not successful. The cycloheximide resistance gene (RIM-C) of Candida maltosa and the URA3 gene of S. cerevisiae were successfully used to transform a prototrophic strain of K. thermotolerans and an Ura-mutant of this yeast isolated in this study, respectively. Transformation was also possible by using intact cells treated with lithiukm salts or thiol compounds. The YRp-type vectors were maintained as plasmids in the transformants under selective conditions. This is the first report of successful transformation of K. thermotolerans.
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