Cloning and expression of Staphylococcus aureus plasmid-mediated quaternary ammonium resistance in Escherichia coli | Zendy

Jan M. Tennent | Zendy; B. R. Lyon | Zendy; Matthew T. Gillespie | Zendy; John W. May | Zendy; Ronald A. Skurray | Zendy

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Cloning and expression of Staphylococcus aureus plasmid-mediated quaternary ammonium resistance in Escherichia coli

Author(s) -

Jan M. Tennent,

B. R. Lyon,

Matthew T. Gillespie,

John W. May,

Ronald A. Skurray

Publication year - 1985

Publication title -

antimicrobial agents and chemotherapy

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 2.07

H-Index - 259

eISSN - 1070-6283

pISSN - 0066-4804

DOI - 10.1128/aac.27.1.79

Subject(s) - plasmid , subcloning , escherichia coli , biology , pbr322 , ethidium bromide , microbiology and biotechnology , kanamycin , restriction enzyme , transposable element , molecular cloning , staphylococcus aureus , dna , genetics , gene , mutant , gene expression , bacteria , antibiotics

The Staphylococcus aureus plasmid pSK1 carries Tn4001, a 4.7-kilobase (kb) transposon which specifies resistance to gentamicin, tobramycin, and kanamycin. In addition, pSK1 mediates resistance to trimethoprim and linked resistance to ethidium bromide (Ebr) and to quaternary ammonium compounds (Qar). Restriction endonuclease analysis of pSK1 and a deleted derivative of pSK1 revealed that the gene(s) responsible for Ebr Qar lies within a 5.2-kb HindIII fragment. This fragment has been cloned into the Escherichia coli plasmid vector pBR322, and transformants of an E. coli K-12 strain exhibited Ebr Qar. Subcloning of the 5.2-kb insert, combined with data from electron microscopic analysis of deleted derivatives of pSK1, located the Ebr Qar determinant(s) on a 2.3-kb segment of pSK1 DNA.

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