Cloning of Schistosoma japonicum Chinese strain 22.6kD membrane-associated protein (Sj-22.6) gene and its overproduction on Escherichia coli

Chin J Biotechnol. 1999;15(1):7-13.

Abstract

A 567bp DNA fragment was amplified from Schistosoma japonicum adult worm mRNA by RT-PCR. Sequence analysis revealed that this fragment contained S. Japonicum Chinese strain membrane-associated protein (Sj-22.6) gene. Then this gene was cloned into the expression vector pGEX-4T, and subsequently expressed in Escherichia coli. The recombinant GST-fusion protein was purified by glutathione agarose affinity chromatography. Its molecular weight was about 48 kD. The yield of expression was around 40 mg/L E.coli culture. The immunological test suggested that the recombinant protein had good antigenity which could make a good basis for the research of its immunological function in Schistosomiasis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Helminth / genetics*
  • Antigens, Helminth / isolation & purification
  • Antigens, Helminth / metabolism
  • Base Sequence
  • Blotting, Western
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Membrane Proteins / genetics*
  • Membrane Proteins / isolation & purification
  • Membrane Proteins / metabolism
  • Mice
  • Molecular Sequence Data
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Schistosoma japonicum / genetics*

Substances

  • 22.6-kDa antigen, Schistosoma japonicum
  • Antigens, Helminth
  • Membrane Proteins
  • Recombinant Proteins