{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T12:33:02Z","timestamp":1753878782977,"version":"3.41.2"},"reference-count":44,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2019,5,7]],"date-time":"2019-05-07T00:00:00Z","timestamp":1557187200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["UNS64750"],"award-info":[{"award-number":["UNS64750"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Helicobacter"],"published-print":{"date-parts":[[2019,8]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p><jats:italic>Helicobacter pylori<\/jats:italic> represents an interesting model of bacterial pathogenesis given that most infections are asymptomatic, while a minority of infections cause severe gastric disease. <jats:italic>H\u00a0pylori<\/jats:italic> strain B128 7.13 is used extensively to understand <jats:italic>H\u00a0pylori<\/jats:italic> pathophysiology. Due to extensive restriction\u2010modification systems, the fact that only some <jats:italic>H\u00a0pylori<\/jats:italic> strains are naturally transformable, the inability of common plasmid and transposon vectors to replicate in this bacterium, as well as the limited number of antibiotic cassettes that are functional in <jats:italic>H\u00a0pylori<\/jats:italic>, there are relatively few genetic tools for the mutagenesis of this bacterium.<\/jats:p><\/jats:sec><jats:sec><jats:title>Materials and Methods<\/jats:title><jats:p>Here, we use PacBio and Illumina sequencing to reveal the complete genome sequence of <jats:italic>H\u00a0pylori<\/jats:italic> B128 7.13. Furthermore, we describe a system to generate markerless and scarless mutations on the <jats:italic>H\u00a0pylori<\/jats:italic> chromosome using the counter\u2010selection marker, galactokinase from <jats:italic>Escherichia coli<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>We show that this mutagenesis strategy can be used to generate in\u2010frame insertions, gene deletions, and multiple independent mutations in B128 7.13. Using the closed genome as a reference, we also report the absence of second site chromosomal mutations and\/or rearrangements in our mutagenized strains. We compare the genome sequence of <jats:italic>H\u00a0pylori<\/jats:italic> B128 7.13 with a closely related strain, <jats:italic>H\u00a0pylori<\/jats:italic> B8, and reveal one notable region of difference, which is a 1430\u00a0bp insertion encoding a <jats:italic>H\u00a0pylori\u2010<\/jats:italic>specific DUF874 family protein of unknown function.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>This article reports the closed genome of the important <jats:italic>H\u00a0pylori<\/jats:italic> B128 7.13 strain and a mutagenesis method that can be adopted by researchers as an alternative strategy to generate isogenic mutants of <jats:italic>H\u00a0pylori<\/jats:italic> in order to further our understanding of this bacterium.<\/jats:p><\/jats:sec>","DOI":"10.1111\/hel.12587","type":"journal-article","created":{"date-parts":[[2019,5,7]],"date-time":"2019-05-07T07:09:55Z","timestamp":1557212995000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Complete genome sequence of <i>Helicobacter pylori<\/i> B128 7.13 and a single\u2010step method for the generation of unmarked mutations"],"prefix":"10.1111","volume":"24","author":[{"given":"Emma M.","family":"Dawson","sequence":"first","affiliation":[{"name":"Institute of Microbiology and Infection College of Medical and Dental Sciences University of Birmingham Birmingham UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Karl A.","family":"Dunne","sequence":"additional","affiliation":[{"name":"Institute of Microbiology and Infection College of Medical and Dental Sciences University of Birmingham Birmingham 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