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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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