Cloning of a DNA sequence that complements glutamine auxotrophy in Saccharomyces cerevisiae.
Glutamine (gln) requiring mutants of Saccharomyces cerevisiae have been isolated. They synthesize small amounts of glutamine synthetase (GS), which is more thermolabile than the enzyme from the parental strain. The gln auxotrophy was complemented in transformation experiments using an S. cerevisiae gene library constructed in the plasmid vector YEp13. The transformants were mitotically unstable and synthesized almost tenfold higher amounts of GS than wild-type cells. This activity was as thermoresistant as that from the wild-type strain. A recombinant plasmid was isolated from one of the transformants and partially mapped. Upon reintroduction into the auxotrophic strain, the transformation frequency to gln prototrophy was the same as that for the marker LEU2 gene. The evidence presented suggests that we have cloned the structural gene for GS from S. cerevisiae.
González
A
A
Dávila
G
G
Calva
E
E
D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004271:DNA, Fungal / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005020:Ethyl Methanesulfonate / Q000494:pharmacology; D005796:Genes; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D005838:Genotype; D005974:Glutamate-Ammonia Ligase / Q000235:genetics*; D005973:Glutamine / Q000378:metabolism*; D009154:Mutation; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
D004271:DNA, Fungal; D005973:Glutamine; D005020:Ethyl Methanesulfonate; D004262:DNA Restriction Enzymes; D005974:Glutamate-Ammonia Ligase