Preliminary study on biological characterization of Chlamydia trachomatis plasmid protein p ORF5

Zhong Yu Li, Yi Mou Wu, Qiu Lin Huang, Guang Ming Zhong

Research output: Contribution to journalArticle

Abstract

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 languageEnglish (US)
Pages (from-to)124-128
Number of pages5
JournalChinese Journal of Microbiology and Immunology
Volume31
Issue number2
DOIs
StatePublished - Dec 1 2011

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Keywords

  • Chlamydia trachomatis
  • Plasmid
  • Secreted protein
  • p

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Virology

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