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We have leveraged this technology to delete the coding sequences for a known type III effector. The evidence provided here indicates that CT694\/CTL0063 is a virulence protein involved in chlamydial invasion. Based on our findings, we designate the gene product corresponding to\n            <jats:italic>ct694-ctl0063<\/jats:italic>\n            <jats:underline>t<\/jats:underline>\n            ranslocated\n            <jats:underline>m<\/jats:underline>\n            embrane-associated\n            <jats:underline>e<\/jats:underline>\n            ffector A (TmeA). Deletion of\n            <jats:italic>tmeA<\/jats:italic>\n            did not impact development of intracellular chlamydiae. However, the absence of TmeA manifested as a decrease in infectivity in both tissue culture and murine infection models. The\n            <jats:italic>in vitro<\/jats:italic>\n            defect was reflected by impaired invasion of host cells. TmeA binds human AHNAK, and we demonstrate here that AHNAK is transiently recruited by invading chlamydiae. TmeA, however, is not required for endogenous AHNAK recruitment. TmeA also impairs AHNAK-dependent actin bundling activity. This TmeA-mediated effect likely does not explain impaired invasion displayed by the\n            <jats:italic>tmeA<\/jats:italic>\n            strain of\n            <jats:named-content content-type=\"genus-species\">Chlamydia<\/jats:named-content>\n            , since AHNAK-deficient cells revealed no invasion phenotype. Overall, our data indicate the efficacy of FRAEM and reveal a role of TmeA during chlamydial invasion that manifests independently of effects on AHNAK.\n          <\/jats:p>","DOI":"10.1128\/iai.00640-17","type":"journal-article","created":{"date-parts":[[2017,10,3]],"date-time":"2017-10-03T00:30:23Z","timestamp":1506990623000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":40,"title":["Fluorescence-Reported Allelic Exchange Mutagenesis Reveals a Role for Chlamydia trachomatis TmeA in Invasion That Is Independent of Host AHNAK"],"prefix":"10.1128","volume":"85","author":[{"given":"M. J.","family":"McKuen","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, Florida, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K. 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