title: Molecular cloning of the cDNA encoding an immunodominant antigen of Dirofilaria immitis.
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 kDa antigenic polypeptide product was demonstrated by immunoprecipitation. cDNA was synthesized from mRNA and a lambda 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 consisted 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.
authors.affiliation: Department of Parasitology, Faculty of Medicine, Shinshu University, Asahi, Japan.
authors.forename: S H
authors.initials: SH
authors.lastname: Sun
authors.forename: T
authors.initials: T
authors.lastname: Matsuura
authors.forename: K
authors.initials: K
authors.lastname: Sugane
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000235:genetics*; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004183:Dirofilaria immitis / Q000235:genetics* / Q000276:immunology; D000939:Epitopes; D005260:Female; D005786:Gene Expression Regulation; D015723:Gene Library; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D012313:RNA / Q000235:genetics; D015183:Restriction Mapping
chemical_list: D000947:Antigens, Helminth; D004274:DNA, Recombinant; D000939:Epitopes; D012313:RNA