A lambda gt11 cDNA recombinant that encodes Dirofilaria immitis paramyosin

Mol Biochem Parasitol. 1989 Jun 1;35(1):31-41. doi: 10.1016/0166-6851(89)90139-4.

Abstract

The cDNA synthesized from mRNA of Dirofilaria immitis female adult worms was cloned into the expression vector lambda gt11. Screening the library with a hyperimmune rabbit antiserum raised against adult worm homogenates yielded several antigen positive clones. One of these clones, lambda cDi2, 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-lambda cDi2 antibodies reacted with a 97-kDa protein on Western transfers of adult D. immitis antigen extracts that were reduced with beta-mercaptoethanol. The whole rabbit anti-D. immitis adult antiserum depleted of anti-lambda cDi2 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 lambda cDi2 showed some similarity to nematode myosin, and gave a stretch of 82 amino acids that is 91.5% identical to Caenorhabditis elegans paramyosin: thus, lambda cDi2 encodes D. immitis paramyosin.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Helminth / immunology
  • Antibodies, Monoclonal / immunology
  • Antigens, Helminth / genetics*
  • Antigens, Helminth / immunology
  • Bacteriophage lambda / genetics
  • Base Sequence
  • Cloning, Molecular / methods
  • Cross Reactions
  • DNA, Recombinant
  • Dirofilaria / genetics*
  • Dirofilaria / growth & development
  • Female
  • Filarioidea / genetics*
  • Immunoblotting
  • Lysogeny
  • Molecular Sequence Data
  • RNA, Messenger / genetics
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics*
  • Recombinant Proteins / immunology
  • Species Specificity
  • Tropomyosin / biosynthesis
  • Tropomyosin / genetics*
  • Tropomyosin / immunology

Substances

  • Antibodies, Helminth
  • Antibodies, Monoclonal
  • Antigens, Helminth
  • DNA, Recombinant
  • RNA, Messenger
  • Recombinant Proteins
  • Tropomyosin

Associated data

  • GENBANK/J04009