Commonly used auxotrophic markers
This table describes some commonly used auxotrophic markers (along with some novel useful markers); it is based on a table in Brachmann 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. Please send e-mail to the curators at SGD at sgd-helpdesk@lists.stanford.edu if you would like to suggest additions or modifications.
Allele | Deleted ORF? | Reverts? | Notes | Molecular Descriptiona | Reference |
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no |
yes |
red colonies |
TGG-to-TGA nonsense change at codon 244; GGA-to-GAA missense change at codon 185 |
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no |
simple point mutation causing a requirement for adenine |
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no |
yes |
ochre mutation |
TTA-to-TTG silent change at codon 9, GAA-to-TAA ochre nonsense change at codon 64, AGA-to-GGA missense change at codon 101, GTT-to-GTC silent change at codon 124, ACG-to-ACA silent change at codon 539 |
Rodney Rothstein, Personal communication to SGD | |
no |
yes |
ochre mutation, red colonies |
G to T transversion at nucleotide 190, changing codon 64 from a Glu to a Stop |
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no |
no |
red colonies |
frameshift (BglII site filled in at position 592) |
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no |
yes |
ochre mutation |
AAA-to-TAA ochre nonsense change at codon 47 |
Rodney Rothstein, Personal communication to SGD | |
yes |
no |
Cold sensitive; high frequency of petite formation, especially during transformation. Note that this deletion damages the PET56 promoter. See Zhang et al. (2003) for a discussion of this issue. |
1 kb deletion (-205 to 835) |
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partial |
no |
chemically constructed deletion; reversion is undetectable (< 10(E-9) revertants in a culture). Causes histidine to be required for growth; allows selection of vectors carrying the HIS3 gene. |
187 bp HindIII-HindIII internal deletion (305 to 492) |
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no |
no |
double mutant |
G deletion at nucleotide 208, G deletion at nucleotide 319 |
Rodney Rothstein, Personal communication to SGD | |
yes |
no |
In the S288C background, the his3 null mutation confers sensitivity to acetic acid. |
complete deletion |
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partial |
no |
EcoRI-ClaI internal deletion (163 to 649, 0.6 kb) |
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no |
no |
double mutant; leu2-3 and leu2-112 are each frameshift mutations; the double mutants revert very infrequently (less than 10(E-8) revertants in a culture) and cause leucine to be required for growth. This allows the selection of vectors carrying the LEU2 gene. |
GTC-to-GTT silent change at codon 56, GTT-to-GCT missense change at codon 69, G insertion at nucleotide 249, G insertion at nucleotide 792, GTT-to-GTC silent change at codon 299, GAC-to-AAC missense change at codon 300 |
Hinnen et al. 1978; Gaber and Culbertson 1982; Meira LB et al., 1995; Rodney Rothstein, Personal communication to SGD Botstein et al. 1979 | |
yes |
no |
In the S288C background, the leu2 null mutation confers sensitivity to acetic acid. |
complete deletion |
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no |
yes |
amber mutation |
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partial |
no |
XhoI-HpaI internal deletion (1813 to 2864, 1.0 kb) |
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yes |
no |
In the S288C background, the lys2 null mutation confers sensitivity to acetic acid. |
complete deletion |
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yes |
no |
cold sensitiveb, weak galactose inducer (deletes GAL3 UAS), removes ARS1, also called trp1-901 |
1.45 kb deletion, EcoRI-EcoRI (-102 to 1352) |
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partial |
no |
cold sensitiveb |
0.6 kb deletion, EcoRI-HindIII (-102 to 513) |
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no |
yes |
amber mutation |
GAG-to-TAG amber nonsense change at codon 83 |
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no |
yes |
cold sensitiveb |
C to T at residue 403 of the coding sequence, changing residue 135 from glutamine to an amber stop codon |
Brian Green and Joachim Li, unpublished results | |
yes |
no |
In the S288C background, the trp1 null mutation confers sensitivity to acetic acid. |
complete deletion |
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no |
yes |
Reverts at low frequency (about 10(E-7) revertants in a culture) and causes tryptophan to be required for growth. |
Allows the selection of vectors carrying the TRP1 gene. |
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yes |
no |
In the S288C background, the trp5 null mutation confers sensitivity to acetic acid. |
complete deletion |
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no |
no |
Ty1 insertion (transcribing left to right) at pos. 121 |
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no |
yes |
G to A transition at residue 701 of the coding sequence, changing residue 234 from glycine to glutamate |
Yan Li, Glenn Manthey, and Adam Bailis, unpublished results | ||
yes |
no |
In the S288C background, the ura3 null mutation confers sensitivity to acetic acid. |
complete deletion |
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yes |
no |
In the S288C background, the ura4 null mutation confers sensitivity to acetic acid. |
complete deletion |
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The alleles listed below are described in Brachmann et al. 1998.
Allele | Deleted ORF? | Reverts? | Notes | Molecular Descriptiona | Reference |
---|---|---|---|---|---|
no |
no |
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yes |
no |
designer deletion |
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yes |
no |
designer deletion |
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yes |
no |
designer deletion |
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yes |
no |
designer deletion |
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aThe sequence coordinates are relative to the first ATG of the selectable marker ORF, in which the A residue is defined as +1.
bAll trp- strains are cold sensitive (Singh and Manney 1974).