Molecular cloning of the cDNA encoding a 42 kDa antigenic polypeptide of Anisakis simplex larvae.
The gene encoding an antigenic polypeptide of Anisakis simplex larvae was studied using recombinant DNA techniques. cDNA synthesized from poly(A)-rich mRNA from A. simplex larvae was ligated into phage vector lambda gtll DNA and packaged in vitro. The phages were propagated on Escherichia coli and a lambda gtll expression library was constructed. A cDNA clone encoding a 42 kDa antigenic polypeptide was selected by immunoscreening of the library and identified by the epitope selection method. A clone containing cDNA for a 42 kDa protein was isolated. The gene encoding this 42 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 42 kDa polypeptide. The antigenic beta-galactosidase fusion protein synthesized by bacteria had no cross-reactivity with other parasite-infected sera.
D000595:Amino Acid Sequence; D000818:Animals; D017163:Anisakis / Q000235:genetics* / Q000276:immunology; D000947:Antigens, Helminth / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis / Q000737:chemistry*; D000939:Epitopes / Q000235:genetics; D005786:Gene Expression Regulation; D015723:Gene Library; D015801:Helminth Proteins / Q000235:genetics*; D007814:Larva / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics
D000947:Antigens, Helminth; D000939:Epitopes; D015801:Helminth Proteins; D012333:RNA, Messenger; D004247:DNA