Cloning of GLN4: an essential gene that encodes glutaminyl-tRNA synthetase in Saccharomyces cerevisiae.
The structural gene for glutaminyl-tRNA synthetase has been isolated from a gene bank of Saccharomyces cerevisiae chromosomal DNA. Cloning was achieved by complementation of a recently described yeast strain that is auxotrophic for glutamine. A multicopy recombinant plasmid with a 5-kilobase-pair genomic insert conferred sixfold elevation in glutaminyl-tRNA synthetase activity and restored a Gln+ phenotype to strains that were Gln- by virtue of a mutant gln4 allele. Subfragments of the 5-kilobase insert directed integration of URA3 to GLN4. Further experiments established that GLN4 is an essential gene that is located on chromosome XV. RNA blots with a GLN4-specific probe detected a single transcript of approximately 2,900 nucleotides.
Ludmerer
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Schimmel
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D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics*; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D005796:Genes; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D005838:Genotype; D009154:Mutation; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D013045:Species Specificity
D004262:DNA Restriction Enzymes; D000604:Amino Acyl-tRNA Synthetases; C020659:glutaminyl-tRNA synthetase