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A positive selection for plasmid loss in Saccharomyces cerevisiae using galactose‐inducible growth inhibitory sequences
Author(s) -
Kawahata Miho,
Amari Shinji,
Nishizawa Yoshinori,
Akada Rinji
Publication year - 1999
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(19990115)15:1<1::aid-yea335>3.0.co;2-9
Subject(s) - plasmid , biology , yeast , galactose , gene , saccharomyces cerevisiae , selection (genetic algorithm) , genetics , biochemistry , artificial intelligence , computer science
Counter‐selections for the loss of introduced plasmid sequences areuseful for gene manipulations in yeast. We have used GAL10 promoter‐mediated overexpression of GIN sequences, whichinhibit the growth of cells, to develop a novel counter‐selectionsystem. Yeast cells carrying a GIN sequence grow normallyon glucose medium but are unable to grow on galactose medium, whereasderivatives that have lost the GIN sequence are able togrow in the presence of galactose. We constructed autonomouslyreplicating, integrating, and disruption plasmids carrying GIN sequences and tested their use to select for loss ofthe plasmid. The results showed that the GIN sequencesprovide a selection for efficient loss of plasmids or integratedconstructs from yeast during growth on galactose medium, indicatingthat this system can be used for plasmid shuffling, gene replacementsand marker gene recycling. This counter‐selection system has wideapplication, because anyGal + strain and a widevariety of marker genes can be used. In addition, counter‐selectionsystems using growth‐inhibitory sequences should be applicable to otheryeasts and possibly to other organisms. Copyright © 1999 John Wiley &Sons, Ltd.