Objective: To localize and characterize the plasmid protein p ORF5 in the Chlamydia trachomatis(Ct) infected cells. Methods: The open reading frame encoding for p ORF5 protein from the Ct plasmid was amplified and cloned into the p GEX-6p vector. The recombinant plasmid p GEX-p ORF5 was transformed into XL1-blue E. coli to express fusion protein with the glutathione-s-transferase (GST). After purified with Glutathione Sepharose 4B beads, the p ORF5 fusion protein was used to immunize mice to make monoclonal and polyclonal antibody. The antibodies were used to localize the endogenous p ORF5 protein and detect the expression pattern in Chlamydia-infected cells using an indirect immunofluorescence assay (IFA). At the same time, ELISA was used to determine whether p ORF5 plasmid protein was expressed and immunogenic during Ct infection in humans. Results: p ORF5 was detected a dominant signal in the cytosol of the Chlamydia-infected cells with a pattern similar to that of anti-CPAF. p ORF5 also appeared in the RBs and EBs in small quantity. Athough pattern was similarly, p ORF5 did not overlap with CPAF. p ORF5 protein was strongly recognized antiserum in an ELISA. Conclusion: The p ORF5 plasmid protein was identified as a secreted protein with good immunogenicity, p ORF5 gene was to express the endogenous target protein during human infection.
|Original language||English (US)|
|Number of pages||5|
|Journal||Chinese Journal of Microbiology and Immunology|
|State||Published - Dec 1 2011|
- Chlamydia trachomatis
- Secreted protein
ASJC Scopus subject areas