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Infection and Immunity, July 1999, p. 3308-3311, Vol. 67, No. 7
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Green Fluorescent Protein as a Marker in Rickettsia typhi Transformation

Jill Michelle Troyer, Suzana Radulovic, and Abdu F. Azad*

Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, Maryland

Received 8 January 1999/Returned for modification 8 February 1999/Accepted 7 April 1999

Transformation of rickettsiae is a recent accomplishment, but utility of this technique is limited due to the paucity of selectable markers suitable for use in this intracellular pathogen. We chose a green fluorescent protein variant optimized for fluorescence under UV lights (GFPUV) as a fluorometric marker and transformed Rickettsia typhi with an rpoB-GFPUV fusion construct. The rickettsiae were subsequently grown in Vero cells, and cultures were screened by PCR and restriction fragment length polymorphism (RFLP) to confirm incorporation of the rpoB-GFPUV construct. Cultures were then analyzed by flow cytometry for detection of GFPUV expression, and transformed R. typhi were isolated in a fluorescence-activated cell sorter. This is the first report of transformation of rickettsiae with a nonrickettsial (GFPUV) gene.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of Maryland School of Medicine, Room 13-009, Bressler Research Building, 655 West Baltimore St., Baltimore, MD 21201. Phone: (410) 706-3335. Fax: (410) 706-0282. E-mail: aazad{at}umaryland.edu.

Infection and Immunity, July 1999, p. 3308-3311, Vol. 67, No. 7
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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