Sets of integrating plasmids and gene disruption cassettes containing improved counter-selection markers designed for repeated use in budding yeast

Yeast
R AkadaY Nishizawa

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 containing auxotrophic markers involving HIS3, TRP1, LEU2, URA3 or ADE2, or a drug-resistant marker PGKp-YAP...Continue Reading

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Citations

Jan 11, 2005·Applied and Environmental Microbiology·Shinji HashimotoRinji Akada
Mar 8, 2013·Annual Review of Food Science and Technology·Hiroshi Kitagaki, Katsuhiko Kitamoto
Apr 20, 2013·Analytical Biochemistry·Jian DongDongguang Xiao
Mar 15, 2016·Metabolic Engineering·Steve SwinnenElke Nevoigt
Mar 16, 2011·Chembiochem : a European Journal of Chemical Biology·Laura M Wingler, Virginia W Cornish
Dec 30, 2014·BMC Genomics·Suchismita ChandranRay-Yuan Chuang
Mar 23, 2013·Metabolic Engineering·Georg HubmannJohan M Thevelein
Sep 18, 2014·FEMS Yeast Research·Michael S SiddiquiChristina D Smolke
Jul 12, 2002·Yeast
Jan 11, 2017·Yeast·Robert Gnügge, Fabian Rudolf
Apr 11, 2017·The Journal of General and Applied Microbiology·Kenshiro Nihei, Tsutomu Kishi
Apr 8, 2018·Current Genetics·Inga SoreanuAmir Aharoni
Nov 15, 2003·BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy·Michael FetchkoIgor Stagljar
May 21, 2009·Applied Microbiology and Biotechnology·Shohei KanekoAkihiko Kondo
Dec 21, 2010·Molecular Biotechnology·Olga A SofyanovichVsevolod A Serebryanyy
Sep 27, 2018·Current Genetics·Neil R AdamesJean Peccoud
Apr 19, 2019·Nucleic Acids Research·Sachiko HayashiTohru Yoshihisa
Nov 9, 2019·Microbial Cell Factories·Hefeng ChenShuang Li
Oct 20, 2005·Journal of Bioscience and Bioengineering·Rinji Akada
Oct 27, 2021·ACS Synthetic Biology·Sang-Woo LeeMorten Otto Alexander Sommer

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