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Infection and Immunity, July 2006, p. 3874-3879, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.01957-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Deletion of znuA Virulence Factor Attenuates Brucella abortus and Confers Protection against Wild-Type Challenge

Veterinary Molecular Biology, Montana State University, Bozeman, Montana 59717-3610

Received 1 December 2005/ Returned for modification 26 January 2006/ Accepted 18 April 2006

znuA is known to be an important factor for survival and normal growth under low Zn²⁺ concentrations for Escherichia coli, Haemophilus spp., Neisseria gonorrhoeae, and Pasteurella multocida. We hypothesized that the znuA gene present in Brucella melitensis 16 M would be similar to znuA in B. abortus and questioned whether it may also be an important factor for growth and virulence of Brucella abortus. Using the B. melitensis 16 M genome sequence, primers were designed to construct a B. abortus deletion mutant. A znuA knockout mutation in B. abortus 2308 (znuA) was constructed and found to be lethal in low-Zn²⁺ medium. When used to infect macrophages, znuA B. abortus showed minimal growth. Further study with znuA B. abortus showed that its virulence in BALB/c mice was attenuated, and most of the bacteria were cleared from the spleen within 8 weeks. Protection studies confirmed the znuA mutant as a potential live vaccine, since protection against wild-type B. abortus 2308 challenge was as effective as that obtained with the RB51 or S19 vaccine strain.

Editor: V. J. DiRita

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