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Journal of Bacteriology, April 2001, p. 2219-2225, Vol. 183, No. 7
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.7.2219-2225.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Determination of Wolbachia Genome Size by Pulsed-Field Gel Electrophoresis

Ling V. Sun,1 Jeremy M. Foster,2 George Tzertzinis,2 Midori Ono,1 Claudio Bandi,3 Barton E. Slatko,2 and Scott L. O'Neill1,*

Section of Vector Biology, Department of Epidemiology and Public Health, Yale University School of Medicine, New Haven, Connecticut 065201; Molecular Parasitology Division, New England Biolabs, Inc., Beverly, Massachusetts 019152; and Istituto di Patologia Generale Veterinaria, Universita di Milano, 20133 Milan, Italy3

Received 5 October 2000/Accepted 8 January 2001

Genome sizes of six different Wolbachia strains from insect and nematode hosts have been determined by pulsed-field gel electrophoresis of purified DNA both before and after digestion with rare-cutting restriction endonucleases. Enzymes SmaI, ApaI, AscI, and FseI cleaved the studied Wolbachia strains at a small number of sites and were used for the determination of the genome sizes of wMelPop, wMel, and wMelCS (each 1.36 Mb), wRi (1.66 Mb), wBma (1.1 Mb), and wDim (0.95 Mb). The Wolbachia genomes studied were all much smaller than the genomes of free-living bacteria such as Escherichia coli (4.7 Mb), as is typical for obligate intracellular bacteria. There was considerable genome size variability among Wolbachia strains, especially between the more parasitic A group Wolbachia infections of insects and the mutualistic C and D group infections of nematodes. The studies described here found no evidence for extrachromosomal plasmid DNA in any of the strains examined. They also indicated that the Wolbachia genome is circular.

* Corresponding author. Mailing address: Department of Epidemiology and Public Health, Yale University School of Medicine, New Haven, CT 06520. Phone: (203) 785-3285. Fax: (203) 785-4782. E-mail: Scott.oneill{at}yale.edu.

Journal of Bacteriology, April 2001, p. 2219-2225, Vol. 183, No. 7
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.7.2219-2225.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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