{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T12:59:29Z","timestamp":1792673969294,"version":"4.5.0"},"reference-count":47,"publisher":"American Society for Microbiology","issue":"24","license":[{"start":{"date-parts":[[2018,12,15]],"date-time":"2018-12-15T00:00:00Z","timestamp":1544832000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000060","name":"NIAID","doi-asserted-by":"crossref","award":["AI065530"],"award-info":[{"award-number":["AI065530"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000060","name":"NIAID","doi-asserted-by":"crossref","award":["AI124649"],"award-info":[{"award-number":["AI124649"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Bacteriol"],"published-print":{"date-parts":[[2018,12,15]]},"abstract":"<jats:p><jats:named-content content-type=\"genus-species\">C. trachomatis<\/jats:named-content>infections represent a significant burden to human health. The ability to genetically manipulate<jats:named-content content-type=\"genus-species\">Chlamydia<\/jats:named-content>spp. is overcoming historic confounding barriers that have impeded rapid progress in understanding overall chlamydial pathogenesis. The current state of genetic manipulation in<jats:named-content content-type=\"genus-species\">Chlamydia<\/jats:named-content>spp. requires further development, including mechanisms to generate markerless gene disruption. We leveraged a stepwise Cre-lox approach to excise selection marker genes from a deleted gene locus. We found this process to be efficient, and the removal of extraneous elements resulted in the reversal of a negative polar effect on a downstream gene. This technique facilitates a more direct assessment of gene function and adds to the<jats:named-content content-type=\"genus-species\">Chlamydia<\/jats:named-content>molecular toolbox by facilitating the deletion of genes within operons.<\/jats:p>","DOI":"10.1128\/jb.00479-18","type":"journal-article","created":{"date-parts":[[2018,9,17]],"date-time":"2018-09-17T14:14:47Z","timestamp":1537193687000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":37,"title":["Floxed-Cassette Allelic Exchange Mutagenesis Enables Markerless Gene Deletion in Chlamydia trachomatis and Can Reverse Cassette-Induced Polar Effects"],"prefix":"10.1128","volume":"200","author":[{"given":"G.","family":"Keb","sequence":"first","affiliation":[{"name":"Department of Microbiology, Immunology &amp; Molecular Genetics, University of Kentucky College of Medicine, Lexington, Kentucky, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R.","family":"Hayman","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology &amp; Molecular Genetics, University of Kentucky College of Medicine, Lexington, Kentucky, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K. A.","family":"Fields","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology &amp; Molecular Genetics, University of Kentucky College of Medicine, Lexington, Kentucky, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","first-page":"139","volume-title":"Chlamydia: intracellular biology, pathogenesis, and immunity","author":"Schachter J","year":"1999","unstructured":"Schachter J. 1999. Infection and disease epidemiology, p 139\u2013169. In Stephens RS (ed), Chlamydia: intracellular biology, pathogenesis, and immunity. ASM Press, Washington, DC."},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.femsre.2005.03.002"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro.2016.30"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-micro-092412-155725"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2003.03462.x"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2008.01.003"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1462-5822.2000.00029.x"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2008.11.009"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1007\/82_2017_76"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro.2017.59"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2018.01.002"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2015.03.014"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002258"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0083989"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01817-15"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0057090"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0076743"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01290-13"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2018.00059"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01442-14"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01034-06"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1999.01523.x"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1003954"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2009.06732.x"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0135295"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00640-17"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-virology-100114-054930"},{"key":"e_1_3_3_29_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.02424-07"},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00881-12"},{"key":"e_1_3_3_31_2","doi-asserted-by":"crossref","first-page":"11A.13.11","DOI":"10.1002\/cpmc.31","article-title":"Chlamydia trachomatis transformation and allelic exchange mutagenesis","volume":"45","author":"Mueller KE","year":"2017","unstructured":"Mueller KE, Wolf K, Fields KA. 2017. Chlamydia trachomatis transformation and allelic exchange mutagenesis. Curr Protocol Microbiol 45:11A.13.11\u201311A.13.15.","journal-title":"Curr Protocol Microbiol"},{"key":"e_1_3_3_32_2","first-page":"97","article-title":"Advances in genetic manipulation of obligate intracellular bacterial pathogens","volume":"2","author":"Beare P","year":"2011","unstructured":"Beare P, Sandoz K, Omsland A, Rockey D, Heinzen R. 2011. Advances in genetic manipulation of obligate intracellular bacterial pathogens. Front Cell Infect Microbiol 2:97.","journal-title":"Front Cell Infect Microbiol"},{"key":"e_1_3_3_33_2","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1128\/9781555817329","volume-title":"Intracellular pathogens I: Chlamydiales","author":"Tan M","year":"2012","unstructured":"Tan M. 2012. Temporal gene regulation during the chlamydial developmental cycle, p 149\u2013169. In Tan M, Bavoil P (ed), Intracellular pathogens I: Chlamydiales. ASM Press, Washington, DC."},{"key":"e_1_3_3_34_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkp1032"},{"key":"e_1_3_3_35_2","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2011-12-10-r98"},{"key":"e_1_3_3_36_2","doi-asserted-by":"crossref","first-page":"MDNA3-0014-2014","DOI":"10.1128\/microbiolspec.MDNA3-0014-2014","article-title":"Cre recombinase","volume":"3","author":"Van Duyne GD","year":"2015","unstructured":"Van Duyne GD. 2015. Cre recombinase. Microbiol Spectr 3:MDNA3-0014-2014.","journal-title":"Microbiol Spectr"},{"key":"e_1_3_3_37_2","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1038\/msb.2013.41","article-title":"Generalized bacterial genome editing using mobile group II introns and Cre-lox","volume":"9","author":"Enyeart PJ","year":"2013","unstructured":"Enyeart PJ, Chirieleison SM, Dao MN, Perutka J, Quandt EM, Yao J, Whitt JT, Keatinge\u2010Clay AT, Lambowitz AM, Ellington AD. 2013. Generalized bacterial genome editing using mobile group II introns and Cre-lox. Mol Syst Biol 9:685.","journal-title":"Mol Syst Biol"},{"key":"e_1_3_3_38_2","doi-asserted-by":"publisher","DOI":"10.1093\/dnares\/dsw015"},{"key":"e_1_3_3_39_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00422-09"},{"key":"e_1_3_3_40_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01592-07"},{"key":"e_1_3_3_41_2","doi-asserted-by":"publisher","DOI":"10.1111\/eva.12196"},{"key":"e_1_3_3_42_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2016.09.006"},{"key":"e_1_3_3_43_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.10.7027-7031.2005"},{"key":"e_1_3_3_44_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13104-015-1542-9"},{"key":"e_1_3_3_45_2","doi-asserted-by":"crossref","DOI":"10.1002\/9780471729259.mc11a01s00","article-title":"Cultivation and laboratory maintenance of Chlamydia trachomatis","volume":"11","author":"Scidmore MA","year":"2005","unstructured":"Scidmore MA. 2005. Cultivation and laboratory maintenance of Chlamydia trachomatis. Curr Protocol Microbiol Chapter 11:Unit 11A.1.","journal-title":"Curr Protocol Microbiol"},{"key":"e_1_3_3_46_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00845-06"},{"key":"e_1_3_3_47_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1102229108"},{"key":"e_1_3_3_48_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2009.11.073"}],"container-title":["Journal of Bacteriology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JB.00479-18","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JB.00479-18","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,9,1]],"date-time":"2022-09-01T17:27:10Z","timestamp":1662053230000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/JB.00479-18"}},"subtitle":[],"editor":[{"given":"Thomas J.","family":"Silhavy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2018,12,15]]},"references-count":47,"journal-issue":{"issue":"24","published-print":{"date-parts":[[2018,12,15]]}},"alternative-id":["10.1128\/JB.00479-18"],"URL":"https:\/\/doi.org\/10.1128\/jb.00479-18","relation":{},"ISSN":["0021-9193","1098-5530"],"issn-type":[{"value":"0021-9193","type":"print"},{"value":"1098-5530","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,12,15]]},"assertion":[{"value":"2018-08-07","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2018-09-12","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2018-11-26","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}