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Complete structure of the gene encoding an immunodominant antigen of Dirofilaria immitis and larva-specific synthesis of primary transcript

Published online by Cambridge University Press:  05 June 2009

S. Sun
Affiliation:
Department of Parasitology, Shinshu University School of Medicine, Asahi 3-1-1, Matsumoto City, Nagano Prefecture, Japan Department of Molecular Biology, Hebei Medical College, Shijiazhuang City, China
K. Sugane*
Affiliation:
Department of Parasitology, Shinshu University School of Medicine, Asahi 3-1-1, Matsumoto City, Nagano Prefecture, Japan
*
*Author for correspondence.

Abstract

The complete gene encoding an immunodominant antigen of Dirofilaria immitis was isolated from a Charomid 9–36 genomic DNA library. This genomic DNA clone termed ‘Dg2’ was characterized by restriction mapping, DNA sequencing of the 5′ flanking region, the exon/intron boundaries and the polyadenylation addition site. The Dg2 with 4872 bp in length consisted of five exons interspersed with four introns. These exons reveal a single open reading frame followed by a long 3′ non-coding region of 1383 bp. The open reading frame of 969 bp encodes a polypeptide of 322 amino acids with a molecular weight of 34,400. The ATG translation initiation codon starts 22 nucleotides downstream from the 5′ end of the first exon. The polyadenylation signal sequence, AATAAA, is located at the 3′ end of the last exon. The transcription initiation site was determined by primer extension technique. S1 nuclease mapping analysis demonstrated that the primary transcript derived from Dg2 is synthesized in microfilariae but not in male or female adult worms. The result suggests that the stage-specific expression of Dg2 is regulated at the level of primary transcript.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 1994

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References

Aviv, H. & Leder, P. (1972) Purification of biologically active globin messenger RNA by chromatography of oligothymidylic acid-cellulose. Proceedings of the National Academy of Sciences (USA) 69, 14081412.CrossRefGoogle ScholarPubMed
Benoist, C., Hare, K.O., Breathnach, R. & Chambon, P. (1980) The obalbumin gene-sequence of putative control regions. Nucleic Acids Research 8, 127142.CrossRefGoogle Scholar
Caravelli, J.F., Zaman, M.B. & Bains, M.S. (1981) Human pulmonary dirofilariasis mimicking metastatic disease. Clinical Bulletin 11, 8890.Google ScholarPubMed
Ciferri, F. (1982) Human pulmonary dirofilariasis in the United States: A critical review. American Journal of Tropical Medicine and Hygiene 3, 302308.CrossRefGoogle Scholar
Dayal, Y. & Neafie, R.C. (1975) Human pulmonary dirofilariasis: A case report and review of the literature. American Review of Respiratory Disease 112, 437443.Google ScholarPubMed
Erondu, N.E. & Donelson, J.E. (1990) Characterization of a myosin-like antigen from Onchocerca volvulus. Molecular and Biochemical Parasitology 40, 213224.CrossRefGoogle ScholarPubMed
Hagenbuchle, O., Santer, M. & Steitz, J.A. (1978) Conservation of the primary structure at the 3′ end of the 18S rRNA from eucaryotic cells. Cell 13, 551563.CrossRefGoogle ScholarPubMed
Hedge, E.C. & Ridley, D.S. (1977) Immunofluorescent reactions with microfilariae. 1, Diagnostic evaluation. Transactions of the Royal Society of Tropical Medicine and Hygiene 71, 304307.CrossRefGoogle ScholarPubMed
Jeska, E.L. & Caruso, J.P. (1986) Collection of fertile Ascaris suum eggs. Journal of Parasitology 72, 964965.CrossRefGoogle ScholarPubMed
Kazura, J.W., Cicirello, H. & Forsyth, K. (1986) Differential recognition of a protective filarial antigen by antibodies from humans with bancroftian filariasis. Journal of Clinical Investigation 77, 19851992.CrossRefGoogle ScholarPubMed
Kozak, M. (1986) Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell 44, 283292.CrossRefGoogle ScholarPubMed
Maniatis, T., Fritsch, E.F. & Sambrook, J. (1982) Molecular cloning: a laboratory manual. New York, Cold Spring Harbour Laboratory Press.Google Scholar
Mizrachi, H.H., Lieberman, P.H., Tolui, S.S. & Sun, T. (1989) Pulmonary dirofilariasis: Mimicry of well-differentiated squamous carcinoma. Human Pathology 20, 818819.CrossRefGoogle ScholarPubMed
Mount, S.M. (1982) A catalogue of splice junction sequences. Nucleic Acids Research 10, 459472.CrossRefGoogle ScholarPubMed
Sanger, F., Nicklen, S. & Coulcon, A.R. (1977) DNA sequencing with chain terminating inhibitors. Proceedings of the National Academy of Sciences (USA) 74, 54635467.CrossRefGoogle ScholarPubMed
Spieth, J., Denison, K., Zuker, E. & Blumenthal, T. (1985) The nucleotide sequence of a nematode vitellogenin gene. Nucleic Acids Research 13, 71297138.CrossRefGoogle ScholarPubMed
Sun, S., Matsuura, T. & Sugane, K. (1991) Molecular cloning of the cDNA encoding an immunodominant antigen of Dirofilaria immitis. Journal of Helminthology 65, 149158.CrossRefGoogle ScholarPubMed
Sun, S. & Sugane, K. (1992) Immunodiagnosis of human dirofilariasis by enzyme-linked immunosorbent assay using recombinant DNA-derived fusion protein. Journal of Helminthology 66, 220226.CrossRefGoogle ScholarPubMed
Sun, S., Matsuura, T. & Sugane, K. (1992) Stage specific expression of a developmentally regulated gene in Dirofilaria immitis. Journal of Helminthology 66, 6267.CrossRefGoogle ScholarPubMed
Watson, J.D., Hopkins, N.H., Roberts, J.W., Steitz, J.A. & Weiner, A.M. (1987) Molecular biology of the gene, 374 pp. California, The Benjamin/Cummings Publishing Company, Inc.Google Scholar
Weil, G.J., Malane, M.S., Powers, K.G. & Blair, L.S. (1985) Monoclonal antibodies to parasite antigens found in the serum of Dirofilaria immitis-infected dogs. Journal of Immunology 134, 11851191.CrossRefGoogle ScholarPubMed
Yazdanbakhsh, M. (1990) Molecular biological approaches towards immunodiagnosis of filariasis. Parasitology Today 6, 207208.CrossRefGoogle ScholarPubMed
Yoshimura, H., Akao, N., Kondo, K., Ohnishi, Y., Kitagawa, M. & Kamimura, K. (1980) Human dirofilariasis in Japan: Case report and review of literature. International Journal of Zoonoses 7, 107114.Google ScholarPubMed