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Antimicrobial Agents and Chemotherapy, April 2002, p. 1038-1045, Vol. 46, No. 4
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.4.1038-1045.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Genetic Determinants of Tetracycline Resistance in Vibrio harveyi

Jeanette W. P. Teo,1 Theresa M. C. Tan,2 and Chit Laa Poh1*

Programme in Environmental Microbiology, Department of Microbiology,1 Department of Biochemistry, Faculty of Medicine, National University of Singapore, Singapore2

Received 13 June 2001/ Returned for modification 3 October 2001/ Accepted 13 January 2002

Isolates of Vibrio harveyi, a prawn pathogen, have demonstrated multiple antibiotic resistance to commonly used antimicrobial agents, such as oxytetracycline. In this paper, we describe the cloning and characterization of two tetracycline resistance determinants from V. harveyi strain M3.4L. The first resistance determinant, cloned as a 4,590-bp fragment, was identical to tetA and flanking sequences encoded on transposon Tn10 from Shigella flexneri. The second determinant, cloned as a 3,358-bp fragment in pATJ1, contains two open reading frames, designated tet35 and txr. tet35 encodes a 369-amino-acid protein that was predicted to have nine transmembrane regions. It is a novel protein which has no homology to any other drug resistance protein but has low levels of homology (28%) to Na+/H+ antiporters. Transposon mutagenesis showed that tet35 and txr were required for tetracycline resistance in a heterologous Escherichia coli host. Tetracycline accumulation studies indicate that E. coli carrying tet35 and txr can function as an energy-dependent tetracycline efflux pump but is less efficient than TetA.

* Corresponding author. Mailing address: Department of Microbiology, Faculty of Medicine, National University of Singapore, 5 Science Dr. 2, Singapore 117 597. Singapore. Phone: (65)-8743674. Fax: (65)-7766872. E-mail: micpohcl{at}nus.edu.sg.

Antimicrobial Agents and Chemotherapy, April 2002, p. 1038-1045, Vol. 46, No. 4
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.4.1038-1045.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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