Molecular cloning and functional expression of enolase from Trichinella spiralis.
The cDNA library was constructed from muscle larvae of Trichinella spiralis, and immunoscreened with sera from mice infected with T. spiralis. A cDNA clone, designated as TsENO, encoded 2-phospho-D-glycerate hydro-lyase (enolase) that catalyzed a reversible conversion of 2-phospho-D-glyceric acid (2PGA) to phosphoenolpyruvate (PEP) in the glycolytic pathway. The recombinant TsENO protein was produced in an Escherichia coli expression system. The recombinant full-length TsENO protein had no activity in the conversion of 2PGA to PEP, but gained enolase activity after cutting off the signal peptide from the full-length protein. There was no meaningful difference in the expression level of TsENO gene at three distinct stages of T. spiralis. Also, antibody against the recombinant TsENO protein reacted with crude extract of muscle larvae, but not with the excretory and secretory products.
Nakada
T
T
Department of Parasitology, Gifu University Graduate School of Medicine, Yanagido 1-1, Gifu 501-1194, Japan.
Nagano
I
I
Wu
Z
Z
Takahashi
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018509:DNA, Helminth / Q000032:analysis; D015723:Gene Library; D015801:Helminth Proteins / Q000235:genetics* / Q000378:metabolism; D008018:Life Cycle Stages / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000469:parasitology; D010751:Phosphopyruvate Hydratase / Q000235:genetics* / Q000378:metabolism; D018510:RNA, Helminth / Q000032:analysis; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017160:Trichinella spiralis / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D014235:Trichinellosis / Q000276:immunology
D018509:DNA, Helminth; D015801:Helminth Proteins; D018510:RNA, Helminth; D011994:Recombinant Proteins; D010751:Phosphopyruvate Hydratase