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Cloning of a probe for a previously undescribed enterobacterial tetracycline resistance gene
Author(s) -
Jones C.S.,
Osborne D.J.,
Stanley J.
Publication year - 1992
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.1992.tb00738.x
Subject(s) - subcloning , biology , genetics , bamhi , tetracycline , bglii , microbiology and biotechnology , plasmid , gene , locus (genetics) , homology (biology) , psti , pbr322 , antibiotics
An 8.35 kb Bam HI fragment was cloned from the plasmid R714a. It encoded resistances to chloramphenicol, streptomycin, spectinomycin and tetracycline. Tetracycline resistance was determined by a locus without homology to the known enterobacterial gene classes, TetA‐TetE. Further subcloning of the fragment located an unusual tetracycline resistance gene on a 4.7 kb Bam HI‐ Bgl II fragment. A physical‐genetic map of this fragment indicated that the gene was bisected by a Pst I site. Deletion analysis and insertion mutagenesis were used to define a suitable probe. An intragenic Pst I‐ Ava I fragment of 1.2 kb was identified, and used as a non‐radioactive probe, being specific for this previously undescribed enterobacterial Tet gene.