[Cloning and cellular localization of pORF8 plasmid protein of Chlamydia trachomatis].

Zhong yu Li, Yi mou Wu, Qiu lin Huang, Sheng mei Su, Zhou Zhou, Chao qun Chen, Guang ming Zhong

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

To clone the plasmid protein pORF8 of Chlamydia trachomatis and localize its expression in Chlamydia-infected cells. pORF8 gene was amplified and cloned into pGEX-6p vector, and the pORF8 fusion protein was expressed in E.coli XL1 Blue. After purification with glutathione-conjugated agarose beads, the pORF8 fusion protein was used to immunize BALB/c mice to generate polyclonal antibodies against pORF8 protein. The antibodies obtained were used to localize the plasmid protein pORF8 in Chlamydia-infected cells with immunofluorescence assay (IFA). The pORF8 gene 744 bp in length was successfully cloned and the GST fusion protein with a relative molecular mass of 54 000 was obtained. The cellular distribution pattern of the plasmid protein pORF8 was similar to that of the major outer membrane protein (MOMP), a known C. trachomatis inclusion body protein, but not to that of chlamydial protease-like activity factor (CPAF, a secreted protein). The plasmid protein pORF8 is localized on the bacterial organism as an inclusion body protein in C. trachomatis-infected cells. The cellular location of pORF8 protein can potentially provide important insights into the pathogenesis of C. trachomatis.

Original languageEnglish (US)
Pages (from-to)1830-1834
Number of pages5
JournalNan fang yi ke da xue xue bao = Journal of Southern Medical University
Volume31
Issue number11
StatePublished - Nov 2011

ASJC Scopus subject areas

  • Medicine(all)

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