Cloning and functional expression of a Schistosoma japonicum cDNA homologous to the enolase gene family

Biochem Biophys Res Commun. 1993 Sep 30;195(3):1211-7. doi: 10.1006/bbrc.1993.2173.

Abstract

A cDNA encoding the complete open reading frame of Schistosoma japonicum enolase has been cloned. The 1494bp cDNA (C30) was isolated from a S. japonicum cDNA expression library immunoscreened with hyperimmune rabbit sera raised against soluble adult S. japonicum proteins. The ORF encodes a protein of 434 amino acids exhibiting 72% identity to human, murine, and rat enolases, and 62% identity to Saccharomyces cerevisiae enolase. The inferred molecular mass of the protein is 47,251 Daltons, similar to that reported for the enolases of other species. In vitro translation of C30 also generated a protein of 47kDa. After subcloning and expression, the recombinant protein was purified by affinity chromatography under non-denaturing conditions and shown to exhibit functional enolase enzymatic activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Molecular Sequence Data
  • Multigene Family / genetics
  • Phosphopyruvate Hydratase / biosynthesis
  • Phosphopyruvate Hydratase / genetics*
  • RNA, Messenger / genetics
  • Recombinant Proteins / biosynthesis
  • Schistosoma japonicum / enzymology
  • Schistosoma japonicum / genetics*
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Species Specificity

Substances

  • RNA, Messenger
  • Recombinant Proteins
  • Phosphopyruvate Hydratase

Associated data

  • GENBANK/D37827
  • GENBANK/L16877
  • GENBANK/L16878
  • GENBANK/L19108
  • GENBANK/L19109
  • GENBANK/L19111
  • GENBANK/L19112
  • GENBANK/L19956
  • GENBANK/L19957
  • GENBANK/L23324