Title | Bacterial expression and purification of recombinant Plasmodium yoelii circumsporozoite protein |
---|
Authors | Stratmann, T., Schmida, S.R., Harperb, J.F. and Kang, A.S. |
---|
Abstract | We report the expression and purification of recombinant rodent malarialPlasmodium yoeliicircumsporozoite surface protein (PyCSP) inEscherichia coli.To facilitate purification of the recombinant protein, the PyCSP was expressed as an amino-terminal fusion protein to glutathioneS-transferase and as a carboxy-terminal fusion protein to a hexahistidyl tag. The expression of the fusion protein was controlled by the inducibletacpromoter. Under optimal conditions the immunoreactive PyCSP represented approximately 0.04% of the total cell lysate. Western blot analysis probing with an anti-PyCSP antibody revealed a wide array of immunoreactive bands. Material isolated by affinity purification on glutathione–Sepharose 4B resin also contained multiple bands indicative of premature termination or carboxyl-terminal degradation. Analysis of protein retained on a nickel nitrilotriacetic acid resin revealed evidence of amino-terminal deleted material. Combining the two mild affinity purifications resulted in isolation of a single immunoreactive protein of approximate molecular weight of 96 kDa. We anticipate that the approach described in this study will facilitate the production of highly purified recombinant proteins. |
---|
Journal | Protein Expression and Purification |
---|
Journal citation | 11 (1), pp. 72-78 |
---|
ISSN | 1046-5928 |
---|
Year | Oct 1997 |
---|
Digital Object Identifier (DOI) | https://doi.org/10.1006/prep.1997.0768 |
---|
Publication dates |
---|
Published | Oct 1997 |
---|