Korean J Parasitol.  2012 Jun;50(2):165-171. 10.3347/kjp.2012.50.2.165.

Expression and Characterization of alpha-Methylacyl CoA Racemase from Anisakis simplex Larvae

Affiliations
  • 1Department of Internal Medicine, Kosin University College of Medicine, Busan 602-703, Korea.
  • 2Department of Pharmacology, Pusan National University School of Medicine, Yangsan 626-870, Korea.
  • 3Department of Parasitology, Pusan National University School of Medicine, Yangsan 626-870, Korea.
  • 4Department of Parasitology, Inje University College of Medicine, Busan 614-735, Korea.
  • 5Department of Parasitology and Genetics, Kosin University College of Medicine, Busan 602-703, Korea. sunnyock@kosin.ac.kr

Abstract

Larval excretory-secretory products of Anisakis simplex are known to cause allergic reactions in humans. A cDNA library of A. simplex 3rd-stage larvae (L3) was immunoscreened with polyclonal rabbit serum raised against A. simplex L3 excretory-secretory products to identify an antigen that elicits the immune response. One cDNA clone, designated as alpha-methylacyl CoA racemase (Amacr) contained a 1,412 bp cDNA transcript with a single open reading frame that encoded 418 amino acids. A. simplex Amacr showed a high degree of homology compared to Amacr orthologs from other species. Amacr mRNA was highly and constitutively expressed regardless of temperature (10-40degrees C) and time (24-48 hr). Immunohistochemical analysis revealed that Amacr was expressed mainly in the ventriculus of A. simplex larvae. The Amacr protein produced in large quantities from the ventriculus is probably responsible for many functions in the development and growth of A. simplex larvae.

Keyword

Anisakis simplex L3; cDNA library; immunoscreening; excretory-secretory product; alpha-methylacyl CoA racemase

MeSH Terms

Amino Acid Sequence
Animals
Anisakis/*enzymology/genetics
Cloning, Molecular
Cluster Analysis
Gene Expression Profiling
Gene Library
Humans
Immunohistochemistry
Larva/enzymology/genetics
Mice
Mice, Inbred ICR
Molecular Sequence Data
Phylogeny
Rabbits
Racemases and Epimerases/genetics/*metabolism
Sequence Homology, Amino Acid
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