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Journal of Bacteriology, November 2002, p. 5926-5934, Vol. 184, No. 21
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.21.5926-5934.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

DNA Inversion on Conjugative Plasmid pVT745

Jinbiao Chen,1 Donald J. Leblanc,2,{dagger} and Dominique M. Galli1*

School of Dentistry, Indiana University, Indianapolis, Indiana 46202,1 Lilly Research Laboratories, Eli Lilly and Co., Indianapolis, Indiana 462852

Received 6 March 2002/ Accepted 31 July 2002

Plasmid pVT745 from Actinobacillus actinomycetemcomitans strain VT745 can be transferred to other A. actinomycetemcomitans strains at a frequency of 10-6. Screening of transconjugants revealed that the DNA of pDMG21A, a pVT745 derivative containing a kanamycin resistance gene, displayed a structural rearrangement after transfer. A 9-kb segment on the plasmid had switched orientation. The inversion was independent of RecA and required the activity of the pVT745-encoded site-specific recombinase. This recombinase, termed Inv, was highly homologous to invertases of the Din family. Two recombination sites of 22 bp, which are arranged in opposite orientation and which function as DNA crossover sequences, were identified on pVT745. One of the sites was located adjacent to the 5' end of the invertase gene, inv. Inversion of the 9-kb segment on pVT745 derivatives has been observed in all A. actinomycetemcomitans strains tested except for the original host, VT745. This would suggest that a host factor that is either inactive or absent in VT745 is required for efficient recombination. Inactivation of the invertase in the donor strain resulted in a 1,000-fold increase in the number of transconjugants upon plasmid transfer. It is proposed that an activated invertase causes the immediate loss of the plasmid in most recipient cells after mating. No biological role has been associated with the invertase as of yet.

* Corresponding author. Mailing address: Indiana University, School of Dentistry, Department of Oral Biology, 1121 W. Michigan St., Indianapolis, IN 46202. Phone: (317) 278-1936. Fax: (317) 278-1411. E-mail: dgalli{at}iupui.edu.

{dagger} Present address: Pharmacia Corporation, Kalamazoo, MI 49001.

Journal of Bacteriology, November 2002, p. 5926-5934, Vol. 184, No. 21
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.21.5926-5934.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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