Protection against cadmium toxicity in yeast by alcohol dehydrogenase.
A cDNA expression library from Schizosaccharomyces pombe was transformed into Saccharomyces cerevisiae to screen for genes capable of conferring cadmium resistance to S. cerevisiae cells. The cDNA library was cloned into the S. cerevisiae expression vector pDB20 which is designed to express cDNAs via the constitutively-expressed promoter of the gene for alcohol dehydrogenase I (ADH1). Terminator and polyadenylation signals are also provided by the ADH1 gene. Cadmium resistant colonies were shown to arise by a recombination event leading to the exchange of the S. pombe DNA with the chromosomal ADH1 gene and a consequent dramatic increase in the ADH1 gene expression due to the high copy number of the plasmid. The overexpression of ADH1 effectively buffered the cells for cadmium ions by formation of Cd-ADH.
Yu
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W
University of Utah Medical Center, School of Medicine, Salt Lake City 84132.
Macreadie
I G
IG
Winge
D R
DR
D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001483:Base Sequence; D002104:Cadmium / Q000302:isolation & purification / Q000378:metabolism / Q000633:toxicity*; D002850:Chromatography, Gel; D015169:Colony Count, Microbial; D004271:DNA, Fungal / Q000235:genetics; D004352:Drug Resistance, Microbial; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics; D014162:Transfection
D004271:DNA, Fungal; D002104:Cadmium; D000426:Alcohol Dehydrogenase