Stratmann, Thomas and Schmida, Stefanie R. and Harperb, Jeffrey F. and Kang, Angray S. (1997) Bacterial expression and purification of recombinant Plasmodium yoelii circumsporozoite protein. Protein Expression and Purification, 11 (1). pp. 72-78. ISSN 1046-5928Full text not available from this repository.
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.
|Subjects:||University of Westminster > Science and Technology > Life Sciences, School of (No longer in use)|
|Depositing User:||Miss Nina Watts|
|Date Deposited:||02 Jan 2008|
|Last Modified:||23 Dec 2009 14:35|
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