Abstract
For genetic manipulation of yeast, numerous selection marker genes have been employed. These include prototrophic markers, markers conferring drug resistance, autoselection markers, and counterselectable markers. This chapter describes the different classes of selection markers and provides a number of examples for different applications.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Chee MK, Haase SB (2012) New and redesigned pRS plasmid shuttle vectors for genetic manipulation of Saccharomyces cerevisiae. G3 (Bethesda) 2:515–526
Brachmann CB, Davies A, Cost GJ et al (1998) Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications. Yeast 14:115–132
Sadowski I, Su T-C, Parent J (2007) Disintegrator vectors for single-copy yeast chromosomal integration. Yeast 24:447–455
Shimoi H, Okuda M, Ito K (2000) Molecular cloning and application of a gene complementing pantothenic acid auxotrophy of sake yeast Kyokai no. 7. J Biosci Bioeng 90:643–647
Sikorski RS, Hieter P (1989) A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122:19–27
Ito-Harashima S, McCusker JH (2004) Positive and negative selection LYS5MX gene replacement cassettes for use in Saccharomyces cerevisiae. Yeast 21:53–61
Cost GJ, Boeke JD (1996) A useful colony colour phenotype associated with the yeast selectable/counter-selectable marker MET15. Yeast 12:939–941
Giersberg M, Degelmann A, Bode R et al (2012) Production of a thermostable alcohol dehydrogenase from Rhodococcus ruber in three different yeast species using the Xplor®2 transformation/expression platform. J Ind Microbiol Biotechnol 39:1385–1396
Goldstein AL, Pan X, McCusker JH (1999) Heterologous URA3MX cassettes for gene replacement in Saccharomyces cerevisiae. Yeast 15:507–511
Gueldener U, Heinisch J, Koehler GJ et al (2002) A second set of loxP marker cassettes for Cre-mediated multiple gene knockouts in budding yeast. Nucleic Acids Res 30:e23
Längle-Rouault F, Jacobs E (1995) A method for performing precise alterations in the yeast genome using a recyclable selectable marker. Nucleic Acids Res 23:3079–3081
Olesen K, Johannesen PF, Hoffmann L et al (2000) The pYC plasmids, a series of cassette-based yeast plasmid vectors providing means of counter-selection. Yeast 16:1035–1043
Wach A, Brachat A, Alberti-Segui C et al (1997) Heterologous HIS3 marker and GFP reporter modules for PCR-targeting in Saccharomyces cerevisiae. Yeast 13:1065–1075
Jakopec V, Walla E, Fleig U (2011) Versatile use of Schizosaccharomyces pombe plasmids in Saccharomyces cerevisiae. FEMS Yeast Res 11:653–655
Ugolini S, Bruschi CV (1996) The red/white colony color assay in the yeast Saccharomyces cerevisiae: epistatic growth advantage of white ade8-18, ade2 cells over red ade2 cells. Curr Genet 30:485–492
Solis-Escalante D, Kuijpers NGA, Bongaerts N et al (2013) amdSYM, a new dominant recyclable marker cassette for Saccharomyces cerevisiae. FEMS Yeast Res 13:126–139
Hartzog PE, Nicholson BP, McCusker JH (2005) Cytosine deaminase MX cassettes as positive/negative selectable markers in Saccharomyces cerevisiae. Yeast 22:789–798
Regenberg B, Hansen J (2000) GAP1, a novel selection and counter-selection marker for multiple gene disruptions in Saccharomyces cerevisiae. Yeast 16:1111–1119
Leite FC, Dos Anjos RS, Basilio AC et al (2013) Construction of integrative plasmids suitable for genetic modification of industrial strains of Saccharomyces cerevisiae. Plasmid 69:114–117
Zhang Y, Wang Z-Y, He X-P et al (2008) New industrial brewing yeast strains with ILV2 disruption and LSD1 expression. Int J Food Microbiol 123:18–24
Kanda K, Ishida T, Hirota R et al (2014) Application of a phosphite dehydrogenase gene as a novel dominant selection marker for yeasts. J Biotechnol 182-183:68–73
Napp SJ, Da Silva NA (1993) Enhancement of cloned gene product synthesis via autoselection in recombinant Saccharomyces cerevisiae. Biotechnol Bioeng 41:801–810
Compagno C, Tura A, Ranzi BM et al (1993) Copy number modulation in an autoselection system for stable plasmid maintenance in Saccharomyces cerevisiae. Biotechnol Prog 9:594–599
Thim L, Hansen MT, Norris K et al (1986) Secretion and processing of insulin precursors in yeast. Proc Natl Acad Sci U S A 83:6766–6770
Song X, Liu Q, Mao J et al (2017) POT1-mediated δ-integration strategy for high-copy, stable expression of heterologous proteins in Saccharomyces cerevisiae. FEMS Yeast Res 17:fox064
Kawasaki GH, Bell L (1999) Stable DNA constructs. U.S. patent 5871957
Unternährer S, Pridmore D, Hinnen A (1991) A new system for amplifying 2 μm plasmid copy number in Saccharomyces cerevisiae. Mol Microbiol 5:1539–1548
Geymonat M, Spanos A, Sedgwick SG (2007) A Saccharomyces cerevisiae autoselection system for optimised recombinant protein expression. Gene 399:120–128
Rech SB, Stateva LI, Oliver SG (1992) Complementation of the Saccharomyces cerevisiae srb1-1 mutation: an autoselection system for stable plasmid maintenance. Curr Genet 21:339–344
Pronk JT (2002) Auxotrophic yeast strains in fundamental and applied research. Appl Environ Microbiol 68:2095–2100
Hottiger T, Kuhla J, Pohlig G et al (1995) 2-μm vectors containing the Saccharomyces cerevisiae metallothionein gene as a selectable marker: excellent stability in complex media, and high-level expression of a recombinant protein from a CUP1-promoter-controlled expression cassette in cis. Yeast 11:1–14
Zhang JG, Liu XY, He XP et al (2011) Improvement of acetic acid tolerance and fermentation performance of Saccharomyces cerevisiae by disruption of the FPS1 aquaglyceroporin gene. Biotechnol Lett 33:277–284
Doignon F, Aigle M, Ribereau-Gayon P (1993) Resistance to imidazoles and triazoles in Saccharomyces cerevisiae as a new dominant marker. Plasmid 30:224–233
Ogawa-Mitsuhashi K, Sagane K, Kuromitsu J et al (2009) MPR1 as a novel selection marker in Saccharomyces cerevisiae. Yeast 26:587–593
Park H, Lopez NI, Bakalinsky AT (1999) Use of sulfite resistance in Saccharomyces cerevisiae as a dominant selectable marker. Curr Genet 36:339–344
van den Berg MA, and. Steensma, H.Y. (1997) Expression cassettes for formaldehyde and fluoroacetate resistance, two dominant markers in Saccharomyces cerevisiae. Yeast 13:551–559
Akada R, Shimizu Y, Matsushita Y et al (2002) Use of a YAP1 overexpression cassette conferring specific resistance to cerulenin and cycloheximide as an efficient selectable marker in the yeast Saccharomyces cerevisiae. Yeast 19:17–28
Fukuda K, Watanabe M, Asano K et al (1992) Molecular breeding of a sake yeast with a mutated ARO4 gene which causes both resistance to o-fluoro-DL-phenylalanine and increased production of β-phenethyl alcohol. J Ferment Bioeng 73:366–369
Hashida-Okado T, Ogawa A, Kato I, Takesako K (1998) Transformation system for prototrophic industrial yeasts using the AUR1 gene as a dominant selection marker. FEBS Lett 425:117–122
del Pozo L, Abarca D, Claros MG, Jiménez A (1991) Cycloheximide resistance as a yeast cloning marker. Curr Genet 19:353–358
Bendoni B, Cavalieri D, Casalone E et al (1999) Trifluoroleucine resistance as a dominant molecular marker in transformation of strains of Saccharomyces cerevisiae isolated from wine. FEMS Microbiol Lett 180:229–233
Lacková D, Šubík J (1999) Use of mutated PDR3 gene as a dominant selectable marker in transformation of prototrophic yeast strains. Folia Microbiol 44:171–176
Xie Q, Jiménez A (1996) Molecular cloning of a novel allele of SMR1 which determines sulfometuron methyl resistance in Saccharomyces cerevisiae. FEMS Microbiol Lett 137:165–168
Kunze G, Bode R, Rintala H, Hofemeister J (1989) Heterologous gene expression of the glyphosate resistance marker and its application in yeast transformation. Curr Genet 15:91–98
Hadfield C, Cashmore AM, Meacock PA (1986) An efficient chloramphenicol-resistance marker for Saccharomyces cerevisiae and Escherichia coli. Gene 45:149–158
Vorachek-Warren MK, McCusker JH (2004) DsdA (D-serine deaminase): a new heterologous MX cassette for gene disruption and selection in Saccharomyces cerevisiae. Yeast 21:163–171
Goldstein AL, McCusker JH (1999) Three new dominant drug resistance cassettes for gene disruption in Saccharomyces cerevisiae. Yeast 15:1541–1553
Wach A, Brachat A, Pöhlmann R, Philippsen P (1994) New heterologous modules for classical or PCR-based gene disruptions in Saccharomyces cerevisiae. Yeast 10:1793–1808
Raymond M, Ruetz S, Thomas DY, Gros P (1994) Functional expression of P-glycoprotein in Saccharomyces cerevisiae confers cellular resistance to the immunosuppressive and antifungal agent FK520. Mol Cell Biol 14:277–286
Miyajima A, Miyajima I, Arai K-I, Arai N (1984) Expression of plasmid R388-encoded type II dihydrofolate reductase as a dominant selective marker in Saccharomyces cerevisiae. Mol Cell Biol 4:407–414
Boeke JD, LaCroute F, Fink GR (1984) A positive selection for mutants lacking orotidine-5′-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet 197:345–346
Chattoo BB, Sherman F, Azubalis DA et al (1979) Selection of lys2 mutants of the yeast Saccharomyces cerevisiae by the utilization of α-aminoadipate. Genetics 93:51–65
Toyn JH, Gunyuzlu PL, White WH et al (2000) A counterselection for the tryptophan pathway in yeast: 5-fluoroanthranilic acid resistance. Yeast 16:553–560
Suizu T, Iimura Y, Gomi K et al (1989) L-Canavanine resistance as a positive selectable marker in diploid yeast transformation through integral disruption of the CAN1 gene. Agric Biol Chem 53:431–436
Akada R, Hirosawa I, Kawahata M et al (2002) Sets of integrating plasmids and gene disruption cassettes containing improved counter-selection markers designed for repeated use in budding yeast. Yeast 19:393–402
Liu Q, Liu H, Yang Y et al (2014) Scarless gene deletion using mazF as a new counter-selection marker and an improved deletion cassette assembly method in Saccharomyces cerevisiae. J Gen Appl Microbiol 60:89–93
Alani E, Cao L, Kleckner N (1987) A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains. Genetics 116:541–545
Reid RJ, Lisby M, Rothstein R (2002) Cloning-free genome alterations in Saccharomyces cerevisiae using adaptamer-mediated PCR. Methods Enzymol 350:258–277
Storici F, Lewis LK, Resnick MA (2001) In vivo site-directed mutagenesis using oligonucleotides. Nat Biotechnol 19:773–776
Fairhead C, Llorente B, Denis F et al (1996) New vectors for combinatorial deletions in yeast chromosomes and for gap-repair cloning using ‘split-marker’ recombination. Yeast 12:1439–1457
Storici F, Durham CL, Gordenin DA, Resnick MA (2003) Chromosomal site-specific double-strand breaks are efficiently targeted for repair by oligonucleotides in yeast. Proc Natl Acad Sci U S A 100:14994–14999
Güldener U, Heck S, Fielder T et al (1996) A new efficient gene disruption cassette for repeated use in budding yeast. Nucleic Acids Res 24:2519–2524
Storici F, Coglievina M, Bruschi CV (1999) A 2-μm DNA-based marker recycling system for multiple gene disruption in the yeast Saccharomyces cerevisiae. Yeast 15:271–283
Babazadeh R, Jafari SM, Zackrisson M et al (2011) The Ashbya gossypii EF-1α promoter of the ubiquitously used MX cassettes is toxic to Saccharomyces cerevisiae. FEBS Lett 585:3907–3913
Wang X, Wang Z, Da Silva NA (1996) G418 Selection and stability of cloned genes integrated at chromosomal delta sequences of Saccharomyces cerevisiae. Biotechnol Bioeng 49:45–51
Loison G, Vidal A, Findeli A et al (1989) High-level of expression of a protective antigen of schistosomes in Saccharomyces cerevisiae. Yeast 5:497–507
Erhart E, Hollenberg CP (1983) The presence of a defective LEU2 gene on 2μ DNA recombinant plasmids of Saccharomyces cerevisiae is responsible for curing and high copy number. J Bacteriol 156:625–635
Chen Y, Partow S, Scalcinati G et al (2012) Enhancing the copy number of episomal plasmids in Saccharomyces cerevisiae for improved protein production. FEMS Yeast Res 12:598–607
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Siewers, V. (2022). An Overview on Selection Marker Genes for Transformation of Saccharomyces cerevisiae. In: Mapelli, V., Bettiga, M. (eds) Yeast Metabolic Engineering. Methods in Molecular Biology, vol 2513. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2399-2_1
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2399-2_1
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2398-5
Online ISBN: 978-1-0716-2399-2
eBook Packages: Springer Protocols