DOI: 10.1111/j.1432-1033.1989.tb14648.x. Molecular cloning, primary structure and disruption of the structural gene of aldolase from <i>Saccharomyces cerevisiae</i>

Molecular cloning, primary structure and disruption of the structural gene of aldolase from Saccharomyces cerevisiae

10.1111/j.1432-1033.1989.tb14648.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

A yeast cDNA genetic library in a bacteriophage expression vector was screened using an antiserum reacting with fructose 1,6‐bisphosphate aldolase from Saccharomyces cerevisiae. Radio‐labelled probes of selected immunopositive clones were used for screening of a yeast genomic library. From the genomic clones a yeast/Escherichia coli shuttle plasmid was constructed containing on a 1990‐base‐pair fragment the entire structural gene FBA1 coding for yeast aldolase. The primary structure of the FBA1 gene was determined. An open reading frame comprises 1077 base pairs coding for a protein of 359 amino acids with a predicted molecular mass of 39608 Da. As observed for other strongly expressed yeast genes, codon usage is extremely biased. The 810 base pairs at the 5′ end and the 90 base pairs at the 3′ end of the coding region of the cloned FBA1 gene are sufficient for normal expression and show characteristic elements present in the noncoding sequences of other yeast genes. Aldolase is the major protein in yeast cells transformed with a high‐copy‐number plasmid containing the FBA1 gene. The aldolase gene was disrupted by insertion of the yeast URA3 gene into the coding region of one FBA1 allele in a homozygous diploid ura3 strain. The haploid offsprings with the defective aldolase allele fbal::URA3lack aldolase enzymatic activity and fail to grow in media containing as a carbon source metabolites of only one side of the aldolase reaction.

Bibliography

SCHWELBERGER, H. G., KOHLWEIN, S. D., & PALTAUF, F. (1989). Molecular cloning, primary structure and disruption of the structural gene of aldolase from Saccharomyces cerevisiae. European Journal of Biochemistry, 180(2), 301â308. Portico.

Authors 3

Hubert Gernot SCHWELBERGER (first)
Sepp Dieter KOHLWEIN (additional)
Fritz PALTAUF (additional)

References 38 Referenced 55

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Dates

Type	When
Created	20 years, 5 months ago (March 3, 2005, 9:48 p.m.)
Deposited	1 year, 8 months ago (Nov. 23, 2023, 4:55 p.m.)
Indexed	11 months, 1 week ago (Sept. 14, 2024, 9:39 p.m.)
Issued	36 years, 5 months ago (March 1, 1989)
Published	36 years, 5 months ago (March 1, 1989)
Published Online	20 years, 5 months ago (March 3, 2005)
Published Print	36 years, 5 months ago (March 1, 1989)

Funders 0

None

BibTeX

@article{SCHWELBERGER_1989, title={Molecular cloning, primary structure and disruption of the structural gene of aldolase from Saccharomyces cerevisiae}, volume={180}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1989.tb14648.x}, DOI={10.1111/j.1432-1033.1989.tb14648.x}, number={2}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={SCHWELBERGER, Hubert Gernot and KOHLWEIN, Sepp Dieter and PALTAUF, Fritz}, year={1989}, month=mar, pages={301–308} }

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