Summary
A diploid strain of the yeast Saccharomyces cerevisiae has been constructed that has one copy of the ribosomal protein gene CYH2 completely deleted and replaced with the TRP1 gene using the method of Rothstein (1983). There are only small differences in growth rate and no detectable difference in steady state level of CYH2 mRNA between the diploid that is heterozygous for the CYH2 deletion and the parent diploid with two normal copies of this gene. This suggests that the diploid must partially compensate for the loss of one CYH2 gene. Tetrad dissection shows that haploid spores lacking the CYH2 gene cannot germinate. The lethality of this deletion can be rescued by a CYH2 cDNA on a low copy vector. Haploids which lack the genomic copy of the CYH2 gene, but contain a plasmid copy of the CYH2 cDNA are able to grow normally. These CYH2 deleted yeast haploids should be useful to analyze mutationally altered CYH2 genes and genes homologous to CYH2 from other organisms without interference from a genomic copy.
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References
Abovich N, Gritz L, Tung L, Rosbash M (1985) Mol Cell Biol 5:3429–3435
Dabbs ER (1986) In: Hardesty B, Kramer G (eds) Structure, function and genetics of ribosomes. Springer, Berlin Heidelberg New York Tokyo, pp 733–748
Cheng S-C, Abelson J (1986) Proc Natl Acad Sci USA 83:2387–2391
Fried HM, Warner JR (1982) Nucleic Acids Res 10:3133–3147
Fried HM, Nam HG, Loechel S, Teem J (1985) Mol Cell Biol 5:99–108
Ito A, Fukuda Y, Murata K, Kimura A (1983) J Bacteriol 153:163–168
Kaufer NF, Fried HM, Schwindinger WF, Jasin M, Warner JR (1983) Nucleic Acids Res 11:3123–3135
McKnight GL, McConaughy BL (1983) Proc Natl Acad Sci USA 80:4412–4416
Mortimer RK, Hawthorne DC (1966) Genetics 53:165–173
Pearson NJ, Fried HM, Warner JR (1982) Cell 34:395–403
Rosseau P, Halvorson H (1973) J Bacteriol 113:1289–1295
Rothstein R (1983) Methods Enzymol 101:202–211
Sherman F, Fink GR, Lawrence CW (eds) (1981) Methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, p 119a
Southern EM (1975) J Mol Biol 98:503–517
Stinchcomb DT, Struhl K, Davis RW (1979) Nature 282:29–43
Stocklein W, Piepersberg W (1980) Curr Genet 1:177–183
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Miles, D.J., Donovan, D.M. & Pearson, N.J. Construction and characterization of a haploid strain of Saccharomyces cerevisiae that completely lacks all genomic CYH2 sequences. Curr Genet 14, 325–329 (1988). https://doi.org/10.1007/BF00419989
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DOI: https://doi.org/10.1007/BF00419989