title: A lambda gt11 cDNA recombinant that encodes Dirofilaria immitis paramyosin.
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.
authors.affiliation: Molecular Parasitology Group, New England Biolabs, Beverly, MA 01915.
authors.forename: A G
authors.initials: AG
authors.lastname: Grandea
authors.forename: L K
authors.initials: LK
authors.lastname: Tuyen
authors.forename: N
authors.initials: N
authors.lastname: Asikin
authors.forename: T B
authors.initials: TB
authors.lastname: Davis
authors.forename: M
authors.initials: M
authors.lastname: Philipp
authors.forename: C
authors.initials: C
authors.lastname: Cohen
authors.forename: L A
authors.initials: LA
authors.lastname: McReynolds
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D000947:Antigens, Helminth / Q000235:genetics* / Q000276:immunology; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D003429:Cross Reactions; D004274:DNA, Recombinant; D004182:Dirofilaria / Q000235:genetics* / Q000254:growth & development; D005260:Female; D005370:Filarioidea / Q000235:genetics*; D015151:Immunoblotting; D008242:Lysogeny; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D013045:Species Specificity; D014335:Tropomyosin / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology
chemical_list: D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D004274:DNA, Recombinant; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014335:Tropomyosin