Cloning and Expression of Novel Aminoglycoside Phosphotransferase Genes from Campylobacter and Their Role in the Resistance to Six Aminoglycosides

S Zhao; S Mukherjee; C Li; S B Jones; S Young; P F McDermott

doi:10.1128/AAC.01682-17

Cloning and Expression of Novel Aminoglycoside Phosphotransferase Genes from Campylobacter and Their Role in the Resistance to Six Aminoglycosides

Antimicrob Agents Chemother. 2017 Dec 21;62(1):e01682-17. doi: 10.1128/AAC.01682-17. Print 2018 Jan.

Authors

S Zhao¹, S Mukherjee², C Li², S B Jones², S Young², P F McDermott²

Affiliations

¹ Division of Animal and Food Microbiology, Office of Research, Center for Veterinary Medicine, U.S. Food and Drug Administration, Laurel, Maryland, USA Shaohua.Zhao@fda.hhs.gov.
² Division of Animal and Food Microbiology, Office of Research, Center for Veterinary Medicine, U.S. Food and Drug Administration, Laurel, Maryland, USA.

Abstract

Nine aph genes, including aph(2″)-Ib, aph(2″)-Ic, aph(2″)-Ig, aph(2″)-If, aph(2″)-If1, aph(2″)-If3, aph(2″)-Ih, aac(6')-Ie-aph(2″)-Ia, and aac(6')-Ie-aph(2″)-If2, were previously identified in Campylobacter To measure the contribution of these alleles to aminoglycoside resistance, we cloned nine genes into the pBluescript and expressed them in Escherichia coli DH5α. The nine aph expressed in E. coli showed various levels of resistance to gentamicin, kanamycin, and tobramycin. Three genes, aac(6″)-Ie-aph(2″)-Ia, aph2″-If1, and aph2″-Ig, showed increased MICs to amikacin, and five aph genes were transferrable.

Keywords: Campylobacter; NARMS; cloning and expression; gentamicin resistance.

This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.

Publication types

Research Support, U.S. Gov't, P.H.S.

MeSH terms

Aminoglycosides / pharmacology*
Anti-Bacterial Agents / pharmacology*
Campylobacter / drug effects
Campylobacter / enzymology*
Campylobacter / genetics*
Cloning, Molecular
Conjugation, Genetic
Drug Resistance, Bacterial / genetics*
Escherichia coli / drug effects
Escherichia coli / genetics
Gene Expression Regulation, Bacterial / drug effects
Gene Expression Regulation, Enzymologic / drug effects
Kanamycin Kinase / biosynthesis
Kanamycin Kinase / genetics*
Microbial Sensitivity Tests

Substances

Aminoglycosides
Anti-Bacterial Agents
Kanamycin Kinase