Parameters affecting the frequencies of transformation and co-transformation with synthetic oligonucleotides in yeast

Yeast. 1992 Nov;8(11):935-48. doi: 10.1002/yea.320081104.


Factors influencing the direct transformation of the yeast Saccharomyces cerevisiae with synthetic oligonucleotides were investigated by selecting for cyc1 transformants that contained at least partially functional iso-1-cytochrome c. Approximately 3 x 10(4) transformants, constituting 0.1% of the cells, were obtained by using 1 mg of oligonucleotide in the reaction mixture. Carrier, such as heterogeneous oligonucleotides, enhanced transformation frequencies. Transformation frequencies were dramatically reduced if the oligonucleotides had a large number of mismatches or had terminally located mismatches. Transformation with oligonucleotides, but not with linearized double-strand plasmid, was efficient in a rad52- strain, suggesting that the pathway for transformation with oligonucleotides is different from that with linearized double-strand plasmid. We describe a procedure of co-transformation with two oligonucleotides, one correcting the cyc1 defect of the target allele in the host strain, and the other producing a desired amino acid alteration elsewhere in the iso-1-cytochrome c molecule; approximately 20% of the transformants obtained by co-transformation contained these desired second alterations.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cytochrome c Group / genetics
  • Cytochromes c*
  • DNA / pharmacology
  • DNA Repair / genetics
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed*
  • Nucleic Acid Hybridization
  • Oligodeoxyribonucleotides / chemical synthesis
  • Oligodeoxyribonucleotides / genetics*
  • RNA, Transfer / pharmacology
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins*
  • Transformation, Genetic* / drug effects


  • CYC1 protein, S cerevisiae
  • Cytochrome c Group
  • Oligodeoxyribonucleotides
  • Saccharomyces cerevisiae Proteins
  • Cytochromes c
  • DNA
  • RNA, Transfer