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PMID: 6206472 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Analysis of full-length cDNA clones carrying GAL1 of Saccharomyces cerevisiae: a model system for cDNA expression.

Nucleic acids research ·Vol. 12 ·No. 16 ·1984-08-24 ·Pages 6397-414

Miyajima A, Nakayama N, Miyajima I, Arai N, Okayama H, Arai K

Abstract

A cDNA cloning vector that allows expression in Saccharomyces cerevisiae has been developed using the plasmid primer approach described by Okayama and Berg [Mol. Cell. Biol. 2:161-170(1982)]. The vector contains ARS1 and TRP1 for plasmid maintenance in yeast and the ADC1 or GAL1 promoter and the TRP5 terminator for expression of the cloned cDNA. Using this system, several recombinants with nearly full-length GAL1 cDNA inserts in a cDNA library made with galactose-induced yeast mRNA were identified. By measurement of galactokinase mRNA and its protein, the expression of GAL1 cDNA was shown to be under the control of the promoter placed upstream of the cDNA insert. Nucleotide sequence analysis revealed that the 3'-ends of the GAL1 cDNA inserts were not unique, indicating that polyA tails were added to GAL1 transcripts at multiple sites in the GAL1 gene. Genetic complementation of appropriate yeast mutants permitted the isolation of clones containing the coding sequences for GAL1, HIS3, and LEU2 from the same cDNA library.

MeSH Terms
Base Sequence Cloning, Molecular DNA/analysis DNA Restriction Enzymes Galactokinase/genetics,metabolism Genes Genes, Fungal Genetic Complementation Test Genetic Vectors Models, Genetic Operon Plasmids Poly A/genetics RNA/genetics RNA, Messenger Saccharomyces cerevisiae/genetics
Chemicals
RNA, Messenger Poly A RNA DNA Galactokinase DNA Restriction Enzymes
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Miyajima A
Nakayama N
Miyajima I
Arai N
Okayama H
Arai K
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30 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/12.16.6397
Published
1984-08-24
Pages
6397-414
Language
English
Region
England
NLM ID
0411011
PMCID
PMC320085
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/12.16.6397 PMC Full Text View journal details
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