Localization and characterization of the hypotheti

Authors: ZhongYu Li QiuLin Huang ShengMei Su Zhou Zhou ChaoQun Chen GuangMing Zhong YiMou Wu

Publish Date: 2011/12/16

Volume: 54, Issue: 11, Pages: 1048-1054

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Abstract

The inclusion membrane proteins play potentially important roles in chlamydial biology and pathogenesis Here we localized and characterized the hypothetical protein CT440 in Chlamydia trachomatisinfected cells The open reading frame ORF encoding the CT440 protein from the C trachomatis serovar D genome was cloned into the prokaryotic expression vector pGEX6p and expressed as a glutathioneStransferase GST fusion protein in E coli XL1Blue The CT440 fusion protein was used to immunize mice to raise antigenspecific antibody After verification by Western blot and immunofluorescence assay IFA the specific antibody was used to localize the endogenous CT440 protein and to detect its expression pattern in Chlamydiainfected cells Cytosolic expression of CT440 in HeLa cells was also carried out to evaluate the effect of the CT440 protein on the subsequent chlamydial infection The results showed that the hypothetical protein CT440 was localized in the C trachomatis inclusion membrane and was detectable 12 h after chlamydial infection Expression of CT440 in the cytoplasm did not inhibit the subsequent chlamydial infection In summary we have identified a new inclusion membrane protein that may be an important candidate for understanding C trachomatis pathogenesisThis article is published under license to BioMed Central Ltd Open Access This article is distributed under the terms of the Creative Commons Attribution License which permits any use distribution and reproduction in any medium provided the original authors and source are creditedThis article is published under an open access license Please check the Copyright Information section for details of this license and what reuse is permitted If your intended use exceeds what is permitted by the license or if you are unable to locate the licence and reuse information please contact the Rights and Permissions team

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