Promoter sequences of the putative Anopheles gambiae apyrase confer salivary gland expression in Drosophila melanogaster.
The saliva of blood-feeding arthropods contains an apyrase that facilitates hematophagy by inhibiting the ADP-induced aggregation of the host platelets. We report here the isolation of a salivary gland-specific cDNA encoding a secreted protein that likely represents the Anopheles gambiae apyrase. We describe also two additional members of the apyrase/5'-nucleotidase family. The cDNA corresponding to the AgApyL1 gene encodes a secreted protein that is closely related in sequence to the apyrase of the yellow fever mosquito, Aedes aegypti, and whose expression appears enriched in, but not restricted to, female salivary glands. The AgApyL2 gene was found searching an A. gambiae data base, and its expression is restricted to larval stages. We isolated the gene encoding the presumed A. gambiae apyrase (AgApy) and we tested its putative promoter for the tissue-specific expression of the LacZ gene from Escherichia coli in transgenic Drosophila melanogaster. All the transgenic lines analyzed showed a weak but unambiguous staining of the adult glands, indicating that some of the salivary gland-specific transcriptional regulatory elements are conserved between the malaria mosquito and the fruit fly. The availability of salivary gland-specific promoters may be useful both for studies on vector-parasite interactions and, potentially, for the targeted tissue-specific expression of anti-parasite genes in the mosquito.
Lombardo
F
F
Istituto di Parassitologia, Istituto Pasteur-Fondazione Cenci Bolognetti, Università di Roma "La Sapienza," 00185 Roma, Italy.
Di Cristina
M
M
Spanos
L
L
Louis
C
C
Coluzzi
M
M
Arcá
B
B
D015720:5'-Nucleotidase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D000330:Aedes / Q000201:enzymology / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D000852:Anopheles / Q000201:enzymology / Q000235:genetics*; D001081:Apyrase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004331:Drosophila melanogaster / Q000235:genetics; D017344:Genes, Insect; D008969:Molecular Sequence Data; D005810:Multigene Family; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism; D012469:Salivary Glands / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D015720:5'-Nucleotidase; D001081:Apyrase