title: [Cloning and cellular localization of pORF8 plasmid protein of Chlamydia trachomatis].
abstract: OBJECTIVE
  To clone the plasmid protein pORF8 of Chlamydia trachomatis and localize its expression in Chlamydia-infected cells.
  
  
  METHODS
  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).
  
  
  RESULTS
  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).
  
  
  CONCLUSIONS
  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.
authors: Department of Microbiology and Immunology, University of South China, Hengyang 421001, China. lzhy1023@hotmail.com
authors: Zhong-yu
authors: ZY
authors: Li
authors: Yi-mou
authors: YM
authors: Wu
authors: Qiu-lin
authors: QL
authors: Huang
authors: Sheng-mei
authors: SM
authors: Su
authors: Zhou
authors: Z
authors: Zhou
authors: Chao-qun
authors: CQ
authors: Chen
authors: Guang-ming
authors: GM
authors: Zhong
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D002690:Chlamydia Infections / Q000378:metabolism; D002692:Chlamydia trachomatis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
chemical_list: D000906:Antibodies; D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins