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Single Nucleotide Polymorphisms in Industrial Yeasts
Chris and Jarlath
Introduction
• Industrial yeasts are aneuploid
• Possess multigene families
• Reorganization events
• Selective process?
Hypothesis
• Low selective pressure leads to silent single nucleotide polymorphisms (SNPs) in multigene families that are important to overall fermentation process
Methods
• Identify candidate genes
• Matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF MS)
• Design Primers
Genes Selected for MALDI-TOF MS
• Degradation of starch
• Glucoamylase multigene family
• STA1 (chr.IV)
• STA2 (chr.II)
• STA3 (chr.XIV)
Information Base Score ESequences producing significant alignments: (bits) Value
emb|X02649|SDSTA1 Yeast (S. diastaticus) STA1 5118 0.0gb|M60650|YSCSTA2A S.cerevisiae STA2 5111 0.0gb|M90490|YSCSTA2K Saccharomyces diastaticus 5073 0.0gb|U30626|SCU30626 Saccharomyces cerevisiae 1108 0.0gb|U95022|SCU95022 Saccharomyces cerevisiae Sta3 860 0.0dbj|D00428|YSCSTA1A Yeast glucoamylase (STA1) gene. 628 0.0
Expected Results
• Detection of SNPs
• Are SNPs random or restricted to hot spots?
• Implications for fermentation characteristics?
Conclusions
• Multicopy, multigene families likely to
contain SNPs
• Implications for overall fermentation process
• Engineered industrial yeast strains
Other Applications
• Genome-wide SNP analysis in humans
e.g. BCRA1
• Pharmacogenomics
ReferencesHinchcliffe, E. (1992) Developing new strains of yeast. J. Inst. Brew.98:27-31.
Johnston, J. R. (1994) Molecular genetics of yeast. Oxford University Press, Oxford, UK.
Lambrechts, M.G., Pretorius, I.S., Marmur, J. and Sollitti, P. (1995) The S1, S2 and SGA1 ancestral genes for the STA glucoamylase genes all map to chromosome IX in Saccharomyces cerevisiae. Yeast 30:783-787.
Lyness, C.A., Jones, C.R., and Meaden P.G. (1993) The STA2 and MEL1 genes of Saccharomyces cerevisiae are idiomorphic. Curr Genet. 23:92-94.
Pretorius, I.S., Lambrechts, M.G., and Marmur, J. (1991) The glucoamylase multigene family in Saccharomyces cerevisiae var. diastaticus: an overview.Crit. Rev. Biochem. Mol. Biol. 26:53-76.
Pugh, T.A., and Clancy, M.J. (1990) Differential regulation of STA genes of Saccharomyces cerevisiae. Mol. Gen. Genet. 222:87-96.
Ross, P., Hall, L., Smirnov, I., and Haff, L. (1998) High level multiplex genotyping by MALDI-TOF mass spectrometry. Nat. Biotech. 16:1347-1351.
http://www.proteome.com
http://morgat.ugs.es/microbsem/v1203-96/benitez.html