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Applied and Environmental Microbiology, December 2006, p. 7718-7722, Vol. 72, No. 12
0099-2240/06/$08.00+0     doi:10.1128/AEM.01578-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Can Anopheles gambiae Be Infected with Wolbachia pipientis? Insights from an In Vitro System{triangledown}

Jason L. Rasgon,* Xiaoxia Ren,{dagger} and Michael Petridis{dagger}

The W. Harry Feinstone Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland 21205,1 Johns Hopkins Malaria Research Institute, Baltimore, Maryland 212052

Received 7 July 2006/ Accepted 2 October 2006

Wolbachia pipientis are maternally inherited endosymbionts associated with cytoplasmic incompatibility, a potential mechanism to drive transgenic traits into Anopheles populations for malaria control. W. pipientis infections are common in many mosquito genera but have never been observed in any Anopheles species, leading to the hypothesis that Anopheles mosquitoes are incapable of harboring infection. We used an in vitro system to evaluate the ability of Anopheles gambiae cells to harbor diverse W. pipientis infections. We successfully established W. pipientis infections (strains wRi and wAlbB) in the immunocompetent Anopheles gambiae cell line Sua5B. Infection was confirmed by PCR, antibiotic curing, DNA sequencing, and direct observation using fluorescence in situ hybridization. The infections were maintained at high passage rates for >30 passages. Our results indicate that there is no intrinsic genetic block to W. pipientis infection in A. gambiae cells, suggesting that establishment of in vivo W. pipientis infections in Anopheles mosquitoes may be feasible.

* Corresponding author. Mailing address: The W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Malaria Research Institute, Bloomberg School of Public Health, Johns Hopkins University, E4626, 615 N. Wolfe Street, Baltimore, MD 21205. Phone: (410) 502-2584. Fax: (410) 955-0105. E-mail: jrasgon{at}jhsph.edu.

{triangledown} Published ahead of print on 6 October 2006.

{dagger} X.R. and M.P. contributed equally to this work.

Applied and Environmental Microbiology, December 2006, p. 7718-7722, Vol. 72, No. 12
0099-2240/06/$08.00+0     doi:10.1128/AEM.01578-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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