Molecular analysis of the gene encoding an antigenic polypeptide of Trichinella spiralis infective larvae.
The gene encoding an antigenic polypeptide of Trichinella spiralis infective larvae was studied using recombinant DNA techniques. cDNA synthesized from poly(A)-rich mRNA from T. spiralis infective larvae was ligated into phage vector lambda gt11 DNA and packaged in vitro. The phages were propagated on Escherichia coli and a lambda gt11 expression library was constructed. A cDNA clone encoding a 46 kDa antigenic polypeptide was selected by immunoscreening of the library and identified by the epitope selection method. A clone containing nearly full-length cDNA for a 46 kDa protein was isolated. The gene encoding this 46 kDa antigenic polypeptide was characterized by DNA and RNA blot analysis using the cDNA as a probe. The gene was transcribed to mRNA with approximately 1400 nucleotides and translated to 46 kDa polypeptide. The antigenic polypeptide was excreted/secreted as a 46 kDa native antigen. The antigenic beta-galactosidase fusion protein synthesized by bacteria had no cross-reactivity with other parasite-infected sera.
Department of Parasitology, Faculty of Medicine, Shinshu University, Matsumoto City, Japan.
K
K
Sugane
T
T
Matsuura
D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D000939:Epitopes / Q000235:genetics; D015801:Helminth Proteins / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000276:immunology; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014158:Transcription, Genetic; D014234:Trichinella / Q000235:genetics* / Q000276:immunology
D000947:Antigens, Helminth; D000939:Epitopes; D015801:Helminth Proteins; D010455:Peptides; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D004247:DNA