Strand-specificity in the transformation of yeast with synthetic oligonucleotides.
Cyc1 mutants of the yeast Saccharomyces cerevisiae were directly transformed with both sense and antisense oligonucleotides to examine the involvement of the two genomic DNA strands in transformation. Sense oligonucleotides yielded approximately 20-fold more transformants than antisense oligonucleotides. This differential effect was observed with oligonucleotides designed to make alterations at six different sites along the gene and was independent of the oligonucleotide sequence and length, number of mismatches and the host strain. Competition studies showed that antisense oligonucleotides did not inhibit transformation. Although the mechanism for this strand specificity is unknown, this difference was maintained even when CYC1 transcription was diminished to approximately 2% of the normal level.
Yamamoto
T
T
Department of Biochemistry, University of Rochester School of Medicine and Dentistry, New York 14642.
Moerschell
R P
RP
Wakem
L P
LP
Komar-Panicucci
S
S
Sherman
F
F
D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D003574:Cytochrome c Group / Q000235:genetics; D045304:Cytochromes c; D016373:DNA, Antisense / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D014158:Transcription, Genetic; D014170:Transformation, Genetic
C014059:CYC1 protein, S cerevisiae; D003574:Cytochrome c Group; D016373:DNA, Antisense; D004271:DNA, Fungal; D009838:Oligodeoxyribonucleotides; D029701:Saccharomyces cerevisiae Proteins; D045304:Cytochromes c