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Nucleic Acids Research, 1983, Vol. 11, No. 3 727-736
© 1983

MOLECULAR BIOLOGY

Identiflcation of clones an E.coli tRNAphe gene by suppression of pheylananyl-tRNA synthetase thermosensitive mutants

Joeèl Caillet, Jacqueline A. Plumbridge, Mathias SPringer, Jean Vacher, Christian Delamarche, Richard H. Buckingham and Marianne Grunberg-Manago

Institut de Biologie Physico-Chimique, 13, rue Pierre et Marie Currie 75005 Paris, France

Received November 19, 1982. Accepted January 5, 1983.

Two libraries of cloned E.coli DNA were screened for plasmids which complemented thermosensitive phenylalanyl-tRNA synthetase mutants. Four plasmids were isolated which complemented pheS and pheT thermosen-sitive mutations but which do not carry pheS or pheT, the structural genes for phenylalanyl-tRNA synthetase. All these plasmids increased the intracellular tRNARne concentration. Three plasmids were shown to carry the structural gene for tRNA"ne which we call pheU. By restriction enzyme analysis, DNA blotting and DNA:tRNA hybridization, pheU was localised to a 280 bp fragment within a 5.6 kb PstIrestriction fragment of E.coli DNA.


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