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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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