{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T00:38:56Z","timestamp":1791851936364,"version":"4.3.4"},"reference-count":77,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T00:00:00Z","timestamp":1764288000000},"content-version":"am","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T00:00:00Z","timestamp":1764288000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T00:00:00Z","timestamp":1764288000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["DE025225"],"award-info":[{"award-number":["DE025225"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["AI105362"],"award-info":[{"award-number":["AI105362"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Molecular Oral Microbiology"],"published-print":{"date-parts":[[2026,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Oral spirochetes are among the small group of keystone pathogens contributing to dysregulation of periodontal tissue homeostasis, leading to breakdown of the tissue and bone supporting the teeth in periodontal disease. Of the more than 60 oral\n                    <jats:italic>Treponema<\/jats:italic>\n                    species and phylotypes,\n                    <jats:italic>Treponema denticola<\/jats:italic>\n                    is one of the few that can be grown in culture and the only one in which genetic manipulation is practicable.\n                    <jats:italic>T. denticola<\/jats:italic>\n                    is thus a model organism for studying spirochete behavior, metabolism, and interactions with other microbes and host tissues that are relevant to oral diseases. We recently demonstrated enhanced transformation efficiency using a synthetic shuttle plasmid resistant to\n                    <jats:italic>T. denticola<\/jats:italic>\n                    restriction\u2013modification systems. Here, we report further optimization of the shuttle plasmid system by minimizing its size and by characterizing an array of promoter\u2010gene constructs for plasmid\u2010based genetic complementation, including the first inducible system for controlled expression of potentially toxic plasmid\u2010encoded genes in\n                    <jats:italic>Treponema<\/jats:italic>\n                    . Our results highlight the importance of precise pairing of promoters and genes of interest for obtaining biologically optimal protein expression. This work expands the utility of the\n                    <jats:italic>T. denticola<\/jats:italic>\n                    shuttle plasmid system and will facilitate future studies in the analysis of\n                    <jats:italic>Treponema<\/jats:italic>\n                    physiology and behavior. Rigorous genetic analysis in oral spirochetes has been hampered by the limited utility of available versions of the\n                    <jats:italic>Escherichia coli\u2013T. denticola<\/jats:italic>\n                    shuttle plasmid system. We report expanded characterization, refinement, and minimization of the shuttle plasmid, including relative activity of diverse promoters and the first inducible expression system described for\n                    <jats:italic>T. denticola<\/jats:italic>\n                    . We show that careful customization of the shuttle plasmid for specific applications is crucial for obtaining successful results.\n                  <\/jats:p>","DOI":"10.1111\/omi.70016","type":"journal-article","created":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T12:11:16Z","timestamp":1764331876000},"page":"69-84","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Expanded Functional Characterization and Optimization of Protein Expression in\n                    <i>Treponema denticola<\/i>\n                    Shuttle Plasmids"],"prefix":"10.1111","volume":"41","author":[{"given":"M. Paula","family":"Goetting\u2010Minesky","sequence":"first","affiliation":[{"name":"Department of Biologic and Materials Sciences and Prosthodontics, School of Dentistry University of Michigan  Ann Arbor Michigan USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valentina","family":"Godovikova","sequence":"additional","affiliation":[{"name":"Department of Biologic and Materials Sciences and Prosthodontics, School of Dentistry University of Michigan  Ann Arbor Michigan USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Prakaimuk","family":"Saraithong","sequence":"additional","affiliation":[{"name":"Department of Biologic and Materials Sciences and Prosthodontics, School of Dentistry University of Michigan  Ann Arbor Michigan USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander H.","family":"Rickard","sequence":"additional","affiliation":[{"name":"Department of Epidemiology, School of Public Health University of Michigan  Ann Arbor Michigan USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brigette R.","family":"Crawley","sequence":"additional","affiliation":[{"name":"Department of Biologic and Materials Sciences and Prosthodontics, School of Dentistry University of Michigan  Ann Arbor Michigan USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara M.","family":"Agolli","sequence":"additional","affiliation":[{"name":"Department of Biologic and Materials Sciences and Prosthodontics, School of Dentistry University of Michigan  Ann Arbor Michigan USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reagan L.","family":"Boyce","sequence":"additional","affiliation":[{"name":"Department of Biologic and Materials Sciences and Prosthodontics, School of Dentistry University of Michigan  Ann Arbor Michigan USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Trishna L.","family":"Appaji","sequence":"additional","affiliation":[{"name":"Department of Biologic and Materials Sciences and Prosthodontics, School of Dentistry University of Michigan  Ann Arbor Michigan USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. Christopher","family":"Fenno","sequence":"additional","affiliation":[{"name":"Department of Biologic and Materials Sciences and Prosthodontics, School of Dentistry University of Michigan  Ann Arbor Michigan USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,11,28]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00078-13"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2018.00732"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1902\/jop.1982.53.9.550"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.12.7851-7857.2001"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.07461-11"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00417-11"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.2.1252-1255.2005"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01537-07"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12033-007-0007-3"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1399-302X.1996.tb00196.x"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.67.7.3653-3656.1999"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.70.4.2233-2237.2002"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.plasmid.2008.10.001"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2017.01479"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1293"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00701-10"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01153-18"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.5.2891-2898.2005"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0757.2005.00108.x"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.263889"},{"key":"e_1_2_9_22_1","first-page":"12B.1.1","volume-title":"Current Protocols in Microbiology","author":"Fenno J. C","year":"2005"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.5.1869-1877.1998"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.178.9.2489-2497.1996"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.1998.tb13047.x"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.63.4.1521-1528.1995"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00046-08"},{"key":"e_1_2_9_28_1","unstructured":"Ghosal D. B.Paul L.Troman et\u00a0al.2025. \u201cStructural Remodeling of Flagellar Filament Promotes Biofilm Formation in Oral PathogenTreponema denticola.\u201d."},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01541-15"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00528-18"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2010.07.020"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2021.668287"},{"key":"e_1_2_9_33_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00228-22"},{"key":"e_1_2_9_34_1","doi-asserted-by":"publisher","DOI":"10.1128\/iai.60.5.2058-2065.1992"},{"key":"e_1_2_9_35_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.2041-1014.2012.00663.x"},{"key":"e_1_2_9_36_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.1991.tb04441.x"},{"key":"e_1_2_9_37_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2008.03.009"},{"key":"e_1_2_9_38_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01031-09"},{"key":"e_1_2_9_39_1","doi-asserted-by":"publisher","DOI":"10.1111\/omi.12441"},{"key":"e_1_2_9_40_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00553-19"},{"key":"e_1_2_9_41_1","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms9061276"},{"key":"e_1_2_9_42_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.plasmid.2005.06.002"},{"key":"e_1_2_9_43_1","first-page":"12B.2.1","volume-title":"Current Protocols in Microbiology","author":"Kuramitsu H. K.","year":"2005"},{"key":"e_1_2_9_44_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.03704-15"},{"key":"e_1_2_9_45_1","doi-asserted-by":"publisher","DOI":"10.1128\/genomeA.01076-14"},{"key":"e_1_2_9_46_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.46.9.2727-2734.2002"},{"key":"e_1_2_9_47_1","doi-asserted-by":"publisher","DOI":"10.1186\/1472-6750-11-92"},{"key":"e_1_2_9_48_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.178.12.3664-3667.1996"},{"key":"e_1_2_9_49_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00478-15"},{"key":"e_1_2_9_50_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.181.12.3743-3750.1999"},{"key":"e_1_2_9_51_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-051X.1978.tb01918.x"},{"key":"e_1_2_9_52_1","doi-asserted-by":"publisher","DOI":"10.1006\/meth.2001.1262"},{"key":"e_1_2_9_53_1","first-page":"62","volume-title":"Host\u2010Parasite Interactions in Periodontal Diseases","author":"Loesche W. J.","year":"1982"},{"key":"e_1_2_9_54_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.174.8.2724-2728.1992"},{"key":"e_1_2_9_55_1","first-page":"39","article-title":"Gene Regulation, Two Component Regulatory Systems, and Adaptive Responses in Treponema denticola","volume":"415","author":"Marconi R. T","year":"2018","journal-title":"Current Topics in Microbiology and Immunology"},{"key":"e_1_2_9_56_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.2041-1014.2010.00598.x"},{"key":"e_1_2_9_57_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01544-08"},{"key":"e_1_2_9_58_1","doi-asserted-by":"publisher","DOI":"10.1128\/am.19.2.207-213.1970"},{"key":"e_1_2_9_59_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0765.1980.tb00268.x"},{"key":"e_1_2_9_60_1","doi-asserted-by":"publisher","DOI":"10.1111\/omi.12458"},{"key":"e_1_2_9_61_1","doi-asserted-by":"publisher","DOI":"10.1111\/tbed.12729"},{"key":"e_1_2_9_62_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.183.12.3770-3783.2001"},{"key":"e_1_2_9_63_1","doi-asserted-by":"publisher","DOI":"10.1093\/femspd\/ftab004"},{"key":"e_1_2_9_64_1","doi-asserted-by":"publisher","DOI":"10.1186\/1754-1611-8-5"},{"key":"e_1_2_9_65_1","first-page":"1","article-title":"The Treponema pallidum Outer Membrane","volume":"415","author":"Radolf J. D.","year":"2018","journal-title":"Current Topics in Microbiology and Immunology"},{"key":"e_1_2_9_66_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1009612"},{"key":"e_1_2_9_67_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.187.5.1792-1798.2005"},{"key":"e_1_2_9_68_1","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.25.11.2230-2232.1987"},{"key":"e_1_2_9_69_1","volume-title":"Molecular Cloning: A Laboratory Manual","author":"Sambrook J.","year":"1989"},{"key":"e_1_2_9_70_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00770-19"},{"key":"e_1_2_9_71_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.2041-1014.2010.00578.x"},{"key":"e_1_2_9_72_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0307639101"},{"key":"e_1_2_9_73_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.72.9.5493-5497.2004"},{"key":"e_1_2_9_74_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep25978"},{"key":"e_1_2_9_75_1","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.13596"},{"key":"e_1_2_9_76_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2017.02353"},{"key":"e_1_2_9_77_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01424-08"},{"key":"e_1_2_9_78_1","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(85)90120-9"}],"container-title":["Molecular Oral Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/omi.70016","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/omi.70016","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/omi.70016","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/omi.70016","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T04:54:00Z","timestamp":1773032040000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/omi.70016"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,28]]},"references-count":77,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2026,4]]}},"alternative-id":["10.1111\/omi.70016"],"URL":"https:\/\/doi.org\/10.1111\/omi.70016","archive":["Portico"],"relation":{},"ISSN":["2041-1006","2041-1014"],"issn-type":[{"value":"2041-1006","type":"print"},{"value":"2041-1014","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,28]]},"assertion":[{"value":"2025-10-10","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-11-19","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-11-28","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}