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Molecular cloning of the cDNA encoding an immunodominant antigen of Dirofilaria immitis

Published online by Cambridge University Press:  05 June 2009

Shu-Han Sun ,
Tadashi Matsuura  and
Kazuo Sugane
Shu-Han Sun
Affiliation:
Department of Parasitology, Faculty of Medicine, Shinshu University, Asahi 3-1-1, Matsumoto City, Nagano Prefecture, Japan
Tadashi Matsuura
Affiliation:
Department of Parasitology, Faculty of Medicine, Shinshu University, Asahi 3-1-1, Matsumoto City, Nagano Prefecture, Japan
Kazuo Sugane*
Affiliation:
Department of Parasitology, Faculty of Medicine, Shinshu University, Asahi 3-1-1, Matsumoto City, Nagano Prefecture, Japan
*
*Author for Correspondence
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Abstract

An immunodominant antigen of Dirofilaria immitis was studied using recombinant DNA techniques. The mRNA from D. immitis adult female worms was translated in vitro and a major 34 k Da antigenic polypeptide product was demonstrated by immunoprecipitation. cDNA was synthesized from mRNA and a λ gt11 expression library was constructed and immunoscreened with dirofilariasis positive serum. A positive clone containing a nearly full length cDNA was isolated. The cDNA was 2415 bp in length and consider of a single open reading frame followed by a long 3' non-coding region of 1446 bp. The open reading frame of 969 bp encoded a polypeptide of 322 amino acids with a molecular weight of 34 400. A cDNA fusion protein synthesized by bacteria (Escherichia coli JM109) using the expression vector pGEMEX-1 was identified as an immunodominant antigen by absorption experiments and had no cross-reactivity with sera from patients with other filarial species.

Keywords

Dirofilaria immitiscDNA libraryantigenic polypeptideimmunoscreening
Type
Research Papers
Information
Journal of Helminthology , Volume 65 , Issue 2 , June 1991 , pp. 149 - 158
Copyright
Copyright © Cambridge University Press 1991

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References

REFERENCES

Arasu, P., Philipp, M. & Perler, F. (1987) Recombinant antigens expressed by genomic DNA clones. Experimental Parasitology, 64, 281–291.CrossRefGoogle ScholarPubMed
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 of the United States of America. 69, 1408–1412.CrossRefGoogle ScholarPubMed
Bemrick, W. J. & Sandholm, H. A. (1966) Aedes vexans and other potential mosquito vectors of Dirofilaria immitis in Minnesota. Journal of Parasitology, 54, 762–767.CrossRefGoogle Scholar
Ciferri, F. (1982) Human pulmonary dirofilariasis in the United States; A critical review. American Journal of Tropical Medicine and Hygiene, 31, 302–308.CrossRefGoogle ScholarPubMed
Dayal, Y. & Neafie, R. C. (1975) Human pulmonary dirofilariasis; A case report and review of the literature. American Review of Respiratory Disease, 112, 437–443.Google ScholarPubMed
Feramisco, J. R., Smart, J. E., Burridge, K., Helfman, D. M. & Thomas, G. P. (1982) Coexistence of vinculin and a vinculin-like protein of higher molecular weight in smooth muscle. Journal of Biological Chemistry, 256, 11024–11031.CrossRefGoogle Scholar
Grandea, A. G. III, Tuyen, L. K., Asikin, N., Davis, T. B., Philipp, M., Cohen, C. & McReynolds, L. A. (1989) A λ gt11 cDNA recombinant that encodes Dirofilaria immitis paramyosin. Molecular and Biochemical Parasitology, 35, 31–42.CrossRefGoogle ScholarPubMed
Gubler, U. & Hoffman, B. J. (1983) A simple and very efficient method for generating cDNA libraries. Gene, 25, 263–269.CrossRefGoogle ScholarPubMed
Heijne, G. V. (1986) A new method for predicting signal sequence cleavage sites. Nucleic Acids Research, 14, 4683–4690.CrossRefGoogle Scholar
Henikoff, S. (1984) Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene, 28, 351–359.CrossRefGoogle ScholarPubMed
Huynh, T. V., Young, R. A. & Davis, R. W. (1984) Constructing and screening cDNA libraries in λ gt10 and λ gt11. In; DNA Cloning (editor Glover, D. M.). pp. 49–78. IRL Press; Oxford, UK.Google Scholar
Irving, D. O. & Howell, M. J. (1981) Preparation and in vitro translation of mRNA from Fasciola hepatica. Molecular and Biochemical Parasitology, 4, 337–348.Google Scholar
Kozak, M. (1986) Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell, 44, 283–292.Google Scholar
Limberger, R. J. & McReynolds, L. A. (1990) Filarial paramyosin; cDNA sequences from Dirofilaria immitis and Onchocerca volvulus. Molecular and Biochemical Parasitology, 38, 2710–280.CrossRefGoogle ScholarPubMed
Nilsen, T. W., Maroney, P. A., Goodwin, R. G., Perrine, K. G., Denker, J. A., Nanduri, J. & Kazura, J. W. (1988) Cloning and characterization of a potentially protective antigen in lymphatic filariasis. Proceedings of the National Academy of Sciences of the United States of America, 85, 3604–3607.CrossRefGoogle ScholarPubMed
Pelham, H. R. B. & Jackson, R. J. (1976) An efficient mRNA-dependent translation system From reticulocyte lysates. European Journal of Biochemistry, 67, 247–256.CrossRefGoogle ScholarPubMed
Philipp, M. & Davis, T. B. (1986) Biochemical and immunologic characterization of a major surface antigen of Dirofilaria immitis infective larvae. Journal of Immunology, 136, 2621–2627.CrossRefGoogle Scholar
Sanger, F., Nicklen, S. & Coulson, A. R. (1977) DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences of the United States of America, 74, 5463–5467.CrossRefGoogle ScholarPubMed
Studier, F. W. & MOFFATT, B. A. (1986) Use of bacteriophage 17 RNA polymerase to direct selective high-level expression of cloned genes. Journal of Molecular Biology, 189, 113–130.Google Scholar
Towbin, H., Staehelin, T. & Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gets to nitrocellulose sheets: Procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America, 76, 4350–4354.CrossRefGoogle Scholar
Unnasch, T. R., Gallin, M. Y., Soboslay, P. T., Erttmann, K. D. & Greene, B. M. (1988) Isolation and characterization of expression cDNA clones encoding antigens of Onchocerca volvulus infective larvae. Journal of Clinical Investigation, 82, 262–269.CrossRefGoogle ScholarPubMed
Weinberger, C., Hollengerg, S. M., Ong, E. S., Harmon, J. M., Brown, S. T., Cidlowski, J., Thompson, F. B., Rosenfeld, M. G. & Evans, R. M. (1985) Identification of human glucocorticoid receptor complementary DNA clones by epitope selection. Science, 228, 740–742.Google Scholar
Yanisch-Perron, C., Vieira, J. & Messing, J. (1985) Improved M13 phage cloning vectors and host strains: Nucleotide sequences of the M13 mp 18 and pUCI9 vectors. Gene, 33. 103–119.CrossRefGoogle Scholar
Yoshimura, H., Akao, N., Kondo, K. & Ohnishi, Y. (1980) Human dirofilariasis in Japan: case report and review of literature. International Journal of Zoonoses, 7, 107–114.Google Scholar