ANALYSIS AND TOXIC OVEREXPRESSION IN ESCHERICHIA-COLI OF A STAPHYLOCOCCAL GENE ENCODING ISOLEUCYL-TRANSFER-RNA SYNTHETASE

作者：CHALKER AF^*; WARD JM; FOSBERRY AP; HODGSON JE

来源：Gene, 1994, 141(1): 103-108.

DOI：10.1016/0378-1119(94)90135-X

• MUPIROCIN
• RECOMBINANT DNA
• OLIGODEOXYRIBONUCLEOTIDE HYBRIDIZATION SCREENING
• STAPHYLOCOCCUS AUREUS
• INCLUSION BODIES

We have cloned and sequenced the Staphylococcus aureus Oxford ileS gene which encodes isoleucyl-tRNA synthetase (Ile-RS), the target for the antibiotic mupirocin. The gene was identified by hybridisation to oligodeoxyribonucleotide probes derived from internal IIe-RS amino acid (aa) sequences. The 2754-bp open reading frame encodes a 918-aa protein of 105 kDa which is homologous to other known Ile-RS from Gram(-) bacteria, archaebacteria, yeast and protozoa. Motifs which have been implicated in the functioning of the active site are strongly conserved. The gene was engineered for high-level expression in Escherichia coli. Ile-RS overproduction was toxic to the E. coli host, the magnitude of its observed effects being strain-dependent.

• 出版日期1994-4-8