A λgt11 cDNA recombinant that encodes Dirofilaria immitis paramyosin

Andres G. Grandea, Le Kim Tuyen, Nur Asikin, Theodore B. Davis, Mario Philipp, Carolyn Cohen, Larry A. McReynolds

Research output: Contribution to journalArticlepeer-review

27 Citations (Scopus)

Abstract

The cDNA synthesized from mRNA of Dirofilaria immitis female adult worms was cloned into the expression vector λgt11. Screening the library with a hyperimmune rabbit antiserum raised against adult worm homogenates yielded several antigen positive clones. One of these clones, λcDi2, was recognized by rabbit antisera raised against either D. immitis L-3, adult, Brugia malayi L-3 or Onchocerca volvulus adult worm antigen, as well as by antisera from humans naturally infected with O. volvulus or Wuchereria bancrofti. Affinity-purified anti-λcDi2 antibodies reacted with a 97-kDa protein on Western transfers of adult D. immitis antigen extracts that were reduced with β-mercaptoethanol. The whole rabbit anti-D. immitis adult antiserum depleted of anti-λcDi2 antibodies exhibited decreased reactivity to this 97-kDa band. A monoclonal antibody (IA6) that specifically binds Schistosoma mansoni paramyosin also recognised a 97-kDa protein in D. immitis extracts upon Western transfer. The deduced amino acid sequence of partial DNA sequence from λcDi2 showed some similarity to nematode myosin, and gave a stretch of 82 amino acids that is 91.5% identical to Caenorhabditis elegans paramyosin: thus, λcDi2 encodes D. immitis paramyosin.

Original languageEnglish
Pages (from-to)31-41
Number of pages11
JournalMolecular and Biochemical Parasitology
Volume35
Issue number1
DOIs
Publication statusPublished - 1 Jun 1989

Keywords

  • Affinity-purified antibody
  • Dirofilaria immitis
  • Myosin
  • Paramyosin
  • λgt11 cDNA library

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