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THE NOVEL PLASMODIUM GALLINACEUM SPOROZOITE PROTEIN, PG93, IS PREFERENTIALLY EXPRESSED IN THE NUCLEUS OF OOCYST SPOROZOITES

To study gene expression differences between oocyst and salivary gland sporozoites, cDNA libraries previously constructed from the two sporozoite populations of the avian malaria parasite, Plasmodium gallinaceum, were used in a subtractive hybridization protocol to isolate Pg93, a novel oocyst sporozoite gene. Pg93 encodes a putative ~76 kDa translated protein that was predicted to localize to the nucleus. Transcriptional analysis indicates that Pg93 is preferentially expressed in oocyst sporozoites versus salivary gland sporozoites. Immunolocalization assays confirm both the nuclear prediction and transcriptional analysis, suggesting that Pg93 is a nuclear protein. BLAST sequence analysis indicates that Pg93 represents a novel gene that has significant homology with a Plasmodium falciparum hypothetical protein and translated Plasmodium knowlesi and Plasmodium vivax nucleotide sequences. This is the first characterization of a Plasmodium nuclear protein that shows preferential expression in one sporozoite population as compared with the other population.

Received November 19, 2004. Accepted for publication March 18, 2005.

Acknowledgments: The authors thank Dr. Louis Miller for the sporozoite cDNA libraries and Dr. David Kaslow for the P. gallinaceum genomic library. The whole genome shotgun sequence data for P. gallinaceum were produced by the Pathogen Sequencing Unit at the Wellcome Trust Sanger Institute and can be obtained from http://www.sanger.ac.uk/Projects/P_gallinaceum.

Financial support: This study was supported by the National Institutes of Health (NIH) grants AI 01657, AI 53156, and F32 AI 09731 to B.T.B and a Minority Biomedical Research Support (MBRTI)/ NIH grant (R25 GM56901) and Missouri Alliance for Graduate Education and the Professoriate (MAGEP)/NSF Fellowship to A.N.L. A.A.J. was supported by a Burroughs-Wellcome Fund Molecular Parasitology Award.

^* Address correspondence to Brenda T. Beerntsen, Department of Veterinary Pathobiology, University of Missouri, 201 Connaway Hall, Columbia, MO 65211. E-mail: BeerntsenB{at}missouri.edu

Authors’ addresses: Alexis N. LaCrue and Brenda T. Beerntsen, Department of Veterinary Pathobiology, University of Missouri, 201 Connaway Hall, Columbia, MO 65211, Telephone: 573-882-5033, Fax: 573-884-5414, E-mail: anebfb{at}missouri.edu and BeerntsenB{at}missouri.edu. Anthony A. James, Microbiology & Molecular Genetics and Molecular Biology & Biochemistry, University of California, 3205 McGaugh Hall, Irvine, CA, 92697, Telephone: 949-824-5930, Fax: 949-824-2814, E-mail: aajames{at}uci.edu.

Reprint requests: Brenda T. Beerntsen, Department of Veterinary Pathology, University of Missouri, 201 Connaway Hall, Columbia, MO 65211.