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PMID: 2829192 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Transformation of yeast with synthetic oligonucleotides.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 85 ·No. 2 ·1988-01-00 ·Pages 524-8

Moerschell RP, Tsunasawa S, Sherman F

Abstract

Genomic DNA of the yeast, Saccharomyces cerevisiae, can be conveniently and specifically altered by transforming spheroplasts or lithium acetate-treated cells directly with synthetic oligonucleotides. Altered forms of iso-1-cytochrome c were generated by transforming a cyc1 mutant with oligonucleotides and selecting for at least partially functional revertants; the oligonucleotides contained a sequence that corrected the cyc1 mutation and produced additional alterations at nearby sites. Transformation has been accomplished with oligonucleotides as short as 20 nucleotides and with amounts as low as 100 micrograms. This method of site-directed mutagenesis in vivo has been used to produce alterations in the NH2-terminal region of iso-1-cytochrome c in which the NH2-terminal methionine is excised and the penultimate residue is acetylated.

MeSH Terms
Alleles Amino Acid Sequence Base Sequence Cytochrome c Group/genetics Cytochromes c Genes Genes, Viral Molecular Sequence Data Oligodeoxyribonucleotides/chemical synthesis Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Transformation, Genetic
Chemicals
CYC1 protein, S cerevisiae Cytochrome c Group Oligodeoxyribonucleotides Saccharomyces cerevisiae Proteins Cytochromes c
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Moerschell R P
Department of Biophysics, University of Rochester School of Medicine and Dentistry, NY 14642.
Tsunasawa S
Sherman F
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.85.2.524
Published
1988-01-00
Pages
524-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC279583
Subset
IM
Grants
NIGMS NIH HHS · R01 GM12702 · United States
External Links
PubMed source DOI: 10.1073/pnas.85.2.524 PMC Full Text View journal details
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