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Molecular cloning and characterization of a gene encoding a 13.1 kDa antigenic protein of Naegleria fowleri.

Authors
Shin, HJ; Cho, MS; Jung, SU; Kim, HI; Park, S; Kim, HJ; Im, KI
Citation
The Journal of eukaryotic microbiology, 48(6):713-717, 2001
Journal Title
The Journal of eukaryotic microbiology
ISSN
1066-52341550-7408
Abstract
An antigen-related gene was cloned from a cDNA expression library of Naegleria fowleri by immunoscreening with sera obtained from mice that were either immunized with an amoebic lysate or infected with trophozoites. The coding nucleotide sequence of the cloned gene consisted of 357 bases that were translated into 119 amino acids. This gene was designated as nfa1. The predicted amino acid sequence of Nfa1 protein has two potential glycosylation and three potential phosphorylation sites, and its predicted secondary structure consists of four helices and three corners. The deduced amino acid sequence of Nfa1 protein shares 43% identity with the myohemerythrin (myoHr) protein from a marine annelid, Nereis diversicolor, including 100% identity in conserved regions and iron-binding residues. A phylogenetic tree constructed from amino acid sequences placed the N. fowleri Nfa1 protein outside of a cluster of myoHr proteins from eight invertebrates. A purified recombinant protein that migrated as a 13.1 kDa species in SDS-PAGE was produced. This recombinant protein exhibited a strong immunoreactivity with infected, immune, and anti-Nfal sera. In addition, an anti-Nfa1 serum reacted with an amoeba lysate in immunoblotting analysis. The present nfal gene encoding the myoHr-like protein is the first myoHr gene cloned from protozoa, and the Nfal antigen may be useful in diagnostic studies.
MeSH terms
Amino Acid SequenceAnimalsAntigens, Protozoan/chemistryAntigens, Protozoan/genetics*Antigens, Protozoan/immunology*Base SequenceCloning, Molecular*Gene LibraryHemerythrin/analogs & derivatives*Hemerythrin/chemistryMiceMice, Inbred BALB CMolecular Sequence DataNaegleria fowleri/geneticsNaegleria fowleri/immunology*Protozoan Proteins/chemistryProtozoan Proteins/genetics*Protozoan Proteins/immunology*Recombinant Proteins/immunologySequence Analysis, DNA
PMID
11831780
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Gastroenterology
Journal Papers > School of Medicine / Graduate School of Medicine > Microbiology
AJOU Authors
신, 호준김, 형일박, 선
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