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Sets of Integrating Plasmids and Gene Disruption Cassettes Containing Improved Counter‐selection Markers Designed for Repeated Use in Budding Yeast by Rinji Akada; Isao Hirosawa; Miho Kawahata; Hisashi Hoshida; Yoshinori Nishizawa is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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## Abstract Counter‐selection is a useful gene manipulation technique for repeated gene disruptions, gene shufflings and gene replacements in yeasts. We developed a novel counter‐selection system using a galactose‐inducible growth inhibitory sequence (Kawahata __et al__.1999. __Yeast__ **15**: 1–10). This counter‐selection marker, named __GAL10p–GIN11__, has several advantages over previous counter‐selection markers, i.e. use of an inexpensive galactose medium for counter‐selection, combined use with any transformation markers for gene introduction, and no requirement of specific mutations in the host strains. The __GIN11__ sequence, which is a part of an X‐element of the subtelomeric regions, contained a conserved autonomously replicating sequence, causing the possibility of inefficient chromosomal integration. We isolated __GIN11__ mutants that lost the replication activity but retained the growth‐inhibitory effect when overexpressed. A mutant __GIN11M86__ sequence was selected and fused to the __CUP1__ promoter for the counter‐selection on a copper‐containing medium. The __GALp–GIN11M86__ and the __CUPp–GIN11M86__ were used for constructing sets of integrating plasmids containin

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Author

Rinji Akada; Isao Hirosawa; Miho Kawahata; Hisashi Hoshida; Yoshinori Nishizawa

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); John Wiley & Sons Inc.; [Chichester]; [New York]: John Wiley & Sons, ©1985-; Wiley; Hindawi Limited (ISSN 0749-503X)

Published

2002

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)