Molecular cloning and characterization of an Rcd1-like protein in excretory-secretory products of Trichinella pseudospiralis.
A cDNA library was constructed from muscle larvae of Trichinella pseudospiralis. A cDNA clone, designated as Tp8 contained a cDNA transcript of 1326 bp length with a single open reading frame, which encoded 303 amino acid residues (34,187 Da, estimated molecular mass). The predicted amino acid sequence of the clone had an identity of approximately 60% to the Rcd1 (Required cell differentiation 1) -like proteins among a wide range of organisms. Real-time quantitative polymerase chain reaction results showed that the transcription level of Tp8 gene reached the highest value in adult worms, and that the transcription level in muscle larvae before stichosome formation was higher than in muscle larvae after stichosome formation. The recombinant Tp8 protein migrated at 37 kDa and reacted to antibody against T. pseudospiralis excretory-secretory (E-S) products and sera from mice infected with T. pseudospiralis. An antibody against the Tp8 recombinant protein could stain proteins migrating at approximately 34 kDa (which is the expected size from the sequence) on Western blotting of E-S products from muscle larvae. An immunocytochemical study showed that the Tp8 protein was present within the stichocyte of muscle larvae and adults worms.
Nagano
I
I
Department of Parasitology, Gifu University Graduate School of Medicine, Yanagido 1-1 Gifu, 501-1194 Japan. isao@cc.gifu-u.ac.jp
Wu
Z
Z
Takahashi
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D018509:DNA, Helminth / Q000032:analysis; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism / Q000469:parasitology; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000737:chemistry; D014234:Trichinella / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity*; D014235:Trichinellosis / Q000469:parasitology
D000947:Antigens, Helminth; C468527:CNOT9 protein, human; D018509:DNA, Helminth; D015801:Helminth Proteins; D011994:Recombinant Proteins; D014157:Transcription Factors