Objective    To obtain purified ClpP molecule produced by its prokaryotic expression system.     Methods    Template DNA was isolated from cultured TIGR4 Streptococcus pneumoniae by gene recombination technology in vitro. Complete ClpP open reading frame (ORF) was cloned into pET-32a expression vector after being confirmed by sequencing. Antigen protein expressed was identified by Western blotting

purified through Ni

2+

affinity chromatography and processed by dialysis and final vacuum freeze drying preservation.     Results    The DNA sequence analysis confirms that the cloned ClpP gene was identical to that in GenBank. ClpP fusion protein was proved by Western blot analysis.     Conclusion    A highly expressed recombinant ClpP fusion antigen protein is successfully obtained

which establishes a material basis on explor

ing the function of ClpP protein and the mechanism of preventing infection of  streptococcus pneumoniae by ClpP.