دورية أكاديمية
أعرض في EDS

Genome editing reveals that pSCL4 is required for chromosome linearity in Streptomyces clavuligerus .

التفاصيل البيبلوغرافية
العنوان: Genome editing reveals that pSCL4 is required for chromosome linearity in Streptomyces clavuligerus .
المؤلفون: Gomez-Escribano JP; Department of Molecular Microbiology, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, UK.; Present address: Department of Bioresources for Bioeconomy and Health Research, Leibniz Institute, DSMZ-German Collection of Microorganisms and Cell Cultures, Inhoffenstraße 7B, 38124 Braunschweig, Germany., Algora Gallardo L; Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0RE, UK., Bozhüyük KAJ; Department of Molecular Microbiology, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, UK.; Present address: Molecular Biotechnology, Department of Biosciences, Goethe University Frankfurt, 60438 Frankfurt am Main, Germany., Kendrew SG; Biotechnology and Environmental Shared Service, GlaxoSmithKline, Southdown View Way, Worthing BN14 8QH, UK.; Engineered Biodesign Limited, Cambridge CB1 3SN, UK., Huckle BD; Biotechnology and Environmental Shared Service, GlaxoSmithKline, Southdown View Way, Worthing BN14 8QH, UK., Crowhurst NA; Biotechnology and Environmental Shared Service, GlaxoSmithKline, Southdown View Way, Worthing BN14 8QH, UK., Bibb MJ; Department of Molecular Microbiology, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, UK., Collis AJ; Biotechnology and Environmental Shared Service, GlaxoSmithKline, Southdown View Way, Worthing BN14 8QH, UK., Micklefield J; Department of Chemistry, Manchester Institute for Biotechnology, University of Manchester, 131 Princess Street, Manchester M1 7DN, UK., Herron PR; Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0RE, UK., Wilkinson B; Department of Molecular Microbiology, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, UK.
المصدر: Microbial genomics [Microb Genom] 2021 Nov; Vol. 7 (11).
نوع المنشور: Journal Article; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: Microbiology Society Country of Publication: England NLM ID: 101671820 Publication Model: Print Cited Medium: Internet ISSN: 2057-5858 (Electronic) Linking ISSN: 20575858 NLM ISO Abbreviation: Microb Genom Subsets: MEDLINE
أسماء مطبوعة: Publication: Oct. 2015- : [London] : Microbiology Society
Original Publication: [London] : Society for General Microbiology, [2015]-
مواضيع طبية MeSH: Gene Editing*/methods , Streptomyces*/genetics, Chromosomes ; Plasmids/genetics
مستخلص: Streptomyces clavuligerus is an industrially important actinomycete whose genetic manipulation is limited by low transformation and conjugation efficiencies, low levels of recombination of introduced DNA, and difficulty in obtaining consistent sporulation. We describe the construction and application of versatile vectors for Cas9-mediated genome editing of this strain. To design spacer sequences with confidence, we derived a highly accurate genome assembly for an isolate of the type strain (ATCC 27064). This yielded a chromosome assembly (6.75 Mb) plus assemblies for pSCL4 (1795 kb) and pSCL2 (149 kb). The strain also carries pSCL1 (12 kb), but its small size resulted in only partial sequence coverage. The previously described pSCL3 (444 kb) is not present in this isolate. Using our Cas9 vectors, we cured pSCL4 with high efficiency by targeting the plasmid's parB gene. Five of the resulting pSCL4-cured isolates were characterized and all showed impaired sporulation. Shotgun genome sequencing of each of these derivatives revealed large deletions at the ends of the chromosomes in all of them, and for two clones sufficient sequence data was obtained to show that the chromosome had circularized. Taken together, these data indicate that pSCL4 is essential for the structural stability of the linear chromosome.
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معلومات مُعتمدة: BBS/E/J/000PR9790 United Kingdom BB_ Biotechnology and Biological Sciences Research Council
فهرسة مساهمة: Keywords: Bionano optical mapping; CRISPR-Cas9; PacBio; actinomycetes genomics; next-generation sequencing; plasmid curing
SCR Organism: Streptomyces clavuligerus
تواريخ الأحداث: Date Created: 20211108 Date Completed: 20220404 Latest Revision: 20240405
رمز التحديث: 20240405
مُعرف محوري في PubMed: PMC8743545
DOI: 10.1099/mgen.0.000669
PMID: 34747689
قاعدة البيانات: MEDLINE
الوصف
تدمد:2057-5858
DOI:10.1099/mgen.0.000669