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Molecular cloning of the cDNA encoding a 42 kDa antigenic polypeptide of Anisakis simplex larvae

Published online by Cambridge University Press:  05 June 2009

K. Sugane ,
S. Sun  and
T. Matsuura
K. Sugane
Affiliation:
Department of Parasitology, Shinshu University School of Medicine, Asahi 3-1-1, Matsumoto City, Naguno Prefecture 390, Japan
S. Sun
Affiliation:
Department of Parasitology, Shinshu University School of Medicine, Asahi 3-1-1, Matsumoto City, Naguno Prefecture 390, Japan
T. Matsuura
Affiliation:
Department of Parasitology, Shinshu University School of Medicine, Asahi 3-1-1, Matsumoto City, Naguno Prefecture 390, Japan
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Abstract

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 λ gtll DNA and packaged in vitro. The phages were propagated on Escherichia coli and a λ 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 β-galactosidase fusion protein synthesized by bacteria had no cross-reactivity with other parasite-infected sera.

Keywords

Anisakis simplexfusion proteinepitope selectionλ gtll
Type
Research Article
Information
Journal of Helminthology , Volume 66 , Issue 1 , March 1992 , pp. 25 - 32
Copyright
Copyright © Cambridge University Press 1992

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