A Cotransformation Method To Identify a Restriction-Modification Enzyme That Reduces Conjugation Efficiency in Campylobacter jejuni

dc.contributor.author Zeng, Ximin
dc.contributor.author Wu, Zuowei
dc.contributor.author Zhang, Qijing
dc.contributor.author Zhang, Qijing
dc.contributor.author Lin, Jun
dc.contributor.department Veterinary Microbiology and Preventive Medicine
dc.date 2018-11-29T05:45:48.000
dc.date.accessioned 2020-07-07T05:15:00Z
dc.date.available 2020-07-07T05:15:00Z
dc.date.copyright Mon Jan 01 00:00:00 UTC 2018
dc.date.embargo 2019-05-01
dc.date.issued 2018-12-01
dc.description.abstract <p>Conjugation is an important mechanism for horizontal gene transfer in Campylobacter jejuni, the leading cause of human bacterial gastroenteritis in developed countries. However, to date, the factors that significantly influence conjugation efficiency in Campylobacter spp. are still largely unknown. Given that multiple recombinant loci could independently occur within one recipient cell during natural transformation, the genetic materials from a high-frequency conjugation (HFC) C. jejuni strain may be cotransformed with a selection marker into a low-frequency conjugation (LFC) recipient strain, creating new HFC transformants suitable for the identification of conjugation factors using a comparative genomics approach. To test this, an erythromycin resistance selection marker was created in an HFC C. jejuni strain; subsequently, the DNA of this strain was naturally transformed into NCTC 11168, an LFC C. jejuni strain, leading to the isolation of NCTC 11168-derived HFC transformants. Whole-genome sequencing analysis and subsequent site-directed mutagenesis identified Cj1051c, a putative restriction-modification enzyme (<em>aka</em> CjeI) that could drastically reduce the conjugation efficiency of NCTC 11168 (>5,000-fold). Chromosomal complementation of three diverse HFC C. jejuni strains with CjeI also led to a dramatic reduction in conjugation efficiency (∼1,000-fold). The purified recombinant CjeI could effectively digest the Escherichia coli-derived shuttle vector pRY107. The endonuclease activity of CjeI was abolished upon short heat shock treatment at 50°C, which is consistent with our previous observation that heat shock enhanced conjugation efficiency in C. jejuni. Together, in this study, we successfully developed and utilized a unique cotransformation strategy to identify a restriction-modification enzyme that significantly influences conjugation efficiency in C. jejuni.</p>
dc.description.comments <p>This article is published as Zeng, Ximin, Zuowei Wu, Qijing Zhang, and Jun Lin. "A Cotransformation Method To Identify a Restriction-Modification Enzyme That Reduces Conjugation Efficiency in Campylobacter jejuni." <em>Applied and Environmental Microbiology</em> 84, no. 23 (2018): e02004-18. DOI: <a href="https://dx.doi.org/10.1128/AEM.02004-18" target="_blank">10.1128/AEM.02004-18</a>. Posted with permission.</p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/vmpm_pubs/206/
dc.identifier.articleid 1205
dc.identifier.contextkey 13349913
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath vmpm_pubs/206
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/92317
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/vmpm_pubs/206/2018_ZhangQijing_CotransformationMethod.pdf|||Fri Jan 14 22:26:28 UTC 2022
dc.source.uri 10.1128/AEM.02004-18
dc.subject.disciplines Digestive System Diseases
dc.subject.disciplines Genetics
dc.subject.disciplines Veterinary Preventive Medicine, Epidemiology, and Public Health
dc.subject.keywords Campylobacter jejuni
dc.subject.keywords comparative genomics
dc.subject.keywords conjugation
dc.subject.keywords restriction-modification enzyme
dc.title A Cotransformation Method To Identify a Restriction-Modification Enzyme That Reduces Conjugation Efficiency in Campylobacter jejuni
dc.type article
dc.type.genre article
dspace.entity.type Publication
relation.isAuthorOfPublication 1c6a5dfc-c604-457f-85be-122910db782e
relation.isOrgUnitOfPublication 16f8e472-b1cd-4d8f-b016-09e96dbc4d83
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