{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T01:16:44Z","timestamp":1792804604641,"version":"4.5.0"},"reference-count":40,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2020,9,2]],"date-time":"2020-09-02T00:00:00Z","timestamp":1599004800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Internal Research Funds of the Witten\/Herdecke University","award":["IFF2017-17"],"award-info":[{"award-number":["IFF2017-17"]}],"id":[{"id":"https:\/\/ror.org\/00yq55g44","id-type":"ROR","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["EH 192\/5-3"],"award-info":[{"award-number":["EH 192\/5-3"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Recently an increasing number of new adenovirus types associated with type-dependent pathogenicity have been identified. However, identification of these clinical isolates represents the very first step to characterize novel pathogens. For deeper analyses, these adenoviruses need to be further characterized in basic virology experiments or they could be applied in translational research. To achieve this goal, it is essential to get genetic access and to enable genetic modification of these novel adenovirus genomes (deletion, insertion, and mutation). Here we demonstrate a high-throughput approach to get genetic access to new adenoviruses via homologous recombination. We first defined the cloning conditions regarding homology arm-length and input adenoviral genome amounts. Then we cloned four naturally occurring adenoviruses (Ad70, Ad73, Ad74, and Ad75) into easy-to-manipulate plasmids and genetically modified them by reporter gene insertion. Three recombinant adenoviruses (Ad70, Ad73, and Ad74) containing a reporter cassette were successfully reconstituted. These novel reporter-labeled adenoviruses were further characterized using the inserted luciferase reporter with respect to receptor usage, presence of anti-adenovirus antibodies, and tropism in vitro. The identified receptor usage, the relatively low prevalence of anti-adenovirus antibodies, and the various cancer cell line transduction pattern are important features of these new pathogens providing essential information for their therapeutic application.<\/jats:p>","DOI":"10.3390\/ijms21176370","type":"journal-article","created":{"date-parts":[[2020,9,2]],"date-time":"2020-09-02T09:29:28Z","timestamp":1599038968000},"page":"6370","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["High-Throughput Cloning and Characterization of Emerging Adenovirus Types 70, 73, 74, and 75"],"prefix":"10.3390","volume":"21","author":[{"given":"Wenli","family":"Zhang","sequence":"first","affiliation":[{"name":"Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, Witten\/Herdecke University, 58453 Witten, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kemal","family":"Mese","sequence":"additional","affiliation":[{"name":"Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, Witten\/Herdecke University, 58453 Witten, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2786-7734","authenticated-orcid":false,"given":"Sebastian","family":"Schellhorn","sequence":"additional","affiliation":[{"name":"Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, Witten\/Herdecke University, 58453 Witten, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nora","family":"Bahlmann","sequence":"additional","affiliation":[{"name":"Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, Witten\/Herdecke University, 58453 Witten, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicolas","family":"Mach","sequence":"additional","affiliation":[{"name":"Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, Witten\/Herdecke University, 58453 Witten, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1483-8827","authenticated-orcid":false,"given":"Oskar","family":"Bunz","sequence":"additional","affiliation":[{"name":"Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, Witten\/Herdecke University, 58453 Witten, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akshay","family":"Dhingra","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Virologie, Adenovirus Konsiliarlabor, Medizinische Hochschule, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elias","family":"Hage","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Virologie, Adenovirus Konsiliarlabor, Medizinische Hochschule, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie-Edith","family":"Lafon","sequence":"additional","affiliation":[{"name":"Microbiologie Fondamentale et Pathog\u00e9nicit\u00e9, Universit\u00e9 de Bordeaux, 33076 Bordeaux, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4764-1708","authenticated-orcid":false,"given":"Harald","family":"Wodrich","sequence":"additional","affiliation":[{"name":"Microbiologie Fondamentale et Pathog\u00e9nicit\u00e9, Universit\u00e9 de Bordeaux, 33076 Bordeaux, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Albert","family":"Heim","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Virologie, Adenovirus Konsiliarlabor, Medizinische Hochschule, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anja","family":"Ehrhardt","sequence":"additional","affiliation":[{"name":"Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, Witten\/Herdecke University, 58453 Witten, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,9,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2895","DOI":"10.1099\/vir.0.19497-0","article-title":"Genetic content and evolution of adenoviruses","volume":"84","author":"Davison","year":"2003","journal-title":"J. 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