Home LiteratureArticle Details
PMID: 6324118 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

In vitro generation of specific deletions in DNA cloned in M13 vectors using synthetic oligodeoxyribonucleotides: mutants in the 5'-flanking region of the yeast alcohol dehydrogenase II gene.

Nucleic acids research ·Vol. 12 ·No. 5 ·1984-03-12 ·Pages 2407-19

Chan VL, Smith M

Abstract

Deletion mutants are particularly useful in defining the boundaries of noncoding genetic functions. Such mutants can be precisely generated using synthetic oligodeoxyribonucleotides as mutagens. In this paper we describe the application of this method to recombinant DNA cloned in a phage M13-derived vector. The mutagenic oligodeoxyribonucleotides, 20 and 21 nucleotides in length, were used to delete a tract of 20 dA-dT base-pairs and an adjacent 22 base-pair perfect dyad from the ADR3 locus, the 5'-flanking regulatory region of the ADR2 gene, of Saccharomyces cerevisiae with high efficiency.

MeSH Terms
Alcohol Dehydrogenase Alcohol Oxidoreductases/genetics Base Sequence Chromosome Deletion Cloning, Molecular DNA Restriction Enzymes DNA, Recombinant/metabolism Genes/drug effects Genes, Fungal/drug effects Genetic Vectors Mutation Nucleic Acid Hybridization Oligodeoxyribonucleotides/pharmacology Oligonucleotides/pharmacology Saccharomyces cerevisiae/enzymology,genetics
Chemicals
DNA, Recombinant Oligodeoxyribonucleotides Oligonucleotides Alcohol Oxidoreductases Alcohol Dehydrogenase DNA Restriction Enzymes
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Chan V L
Smith M
References (17)
17 references, click to expand
  1. Cis-dominant regulatory mutations affecting the formation of glucose-repressible alcohol dehydrogenase (ADHII) in Saccharomyces cerevisiae.
    Mol Gen Genet. 1976 Jun 15;145(3):327-33 PMID: 781520
  2. Isolation and characterization of further cis- and trans-acting regulatory elements involved in the synthesis of glucose-repressible alcohol dehydrogenase (ADHII) in Saccharomyces cerevisiae.
    Mol Gen Genet. 1979 Nov;176(3):427-31 PMID: 392242
  3. Use of E. coli polynucleotide phosphorylase for the synthesis of oligodeoxyribonucleotides of defined sequence.
    Methods Enzymol. 1980;65(1):687-701 PMID: 6990191
  4. Directed deletion of a yeast transfer RNA intervening sequence.
    Science. 1980 Sep 19;209(4463):1396-400 PMID: 6997991
  5. Transposable elements associated with constitutive expression of yeast alcohol dehydrogenase II.
    Cell. 1981 Feb;23(2):605-14 PMID: 6258806
  6. A system for shotgun DNA sequencing.
    Nucleic Acids Res. 1981 Jan 24;9(2):309-21 PMID: 6259625
  7. Site-specific mutagenesis using oligodeoxyribonucleotides: isolation of a phenotypically silent phi X174 mutant, with a specific nucleotide deletion, at very high efficiency.
    Gene. 1980 Dec;12(1-2):129-37 PMID: 6452306
  8. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing.
    J Mol Biol. 1980 Oct 25;143(2):161-78 PMID: 6260957
  9. Yeast gene TRP5: structure, function, regulation.
    J Biol Chem. 1982 Feb 10;257(3):1491-500 PMID: 6276387
  10. The nucleotide sequence of the HIS4 region of yeast.
    Gene. 1982 Apr;18(1):47-59 PMID: 7049842
  11. Characterization of a regulatory region upstream of the ADR2 locus of S. cerevisiae.
    Nature. 1982 Dec 23;300(5894):724-8 PMID: 6757760
  12. Oligonucleotide-directed mutagenesis using M13-derived vectors: an efficient and general procedure for the production of point mutations in any fragment of DNA.
    Nucleic Acids Res. 1982 Oct 25;10(20):6487-500 PMID: 6757864
  13. An expandable gene that encodes a Drosophila glue protein is not expressed in variants lacking remote upstream sequences.
    Cell. 1982 Jul;29(3):1041-51 PMID: 6817924
  14. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments.
    Gene. 1982 Oct;19(3):269-76 PMID: 6295880
  15. Nucleotide sequence of the yeast alcohol dehydrogenase II gene.
    J Biol Chem. 1983 Feb 25;258(4):2674-82 PMID: 6337160
  16. Characterization of transposable element-associated mutations that alter yeast alcohol dehydrogenase II expression.
    Mol Cell Biol. 1983 Jan;3(1):20-31 PMID: 6298605
  17. DNA sequences of two yeast promoter-up mutants.
    Nature. 1983 Aug 18-24;304(5927):652-4 PMID: 6348555
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
Published
1984-03-12
Pages
2407-19
Language
English
Region
England
NLM ID
0411011
PMCID
PMC318671
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]