Open AccessIdentiflcation of clones anE.colitRNAphegene by suppression of pheylananyl-tRNA synthetase thermosensitive mutants
Author(s) -
Joeèl Caillet,
Jacqueline Plumbridge,
Mathias Springer,
Jean Vacher,
Christian Delamarche,
Richard H. Buckingham,
Marianne GrunbergManago
Publication year - 1983
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/11.3.727
Subject(s) - biology , plasmid , microbiology and biotechnology , transfer rna , restriction enzyme , gene , mutant , restriction map , dna , psti , structural gene , restriction fragment , southern blot , escherichia coli , genetics , rna
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 thermosensitive mutations but which do not carry pheS or pheT, the structural genes for phenylalanyl-tRNA synthetase. All these plasmids increased the intracellular tRNAPhe concentration. Three plasmids were shown to carry the structural gene for tRNAPhe 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 PstI restriction fragment of E.coli DNA.