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  A Tailored galK Counterselection System for Efficient Markerless Gene Deletion and Chromosomal Tagging in Magnetospirillum gryphiswaldense

Raschdorf, O., Plitzko, J. M., Schüler, D., & Müller, F. D. (2014). A Tailored galK Counterselection System for Efficient Markerless Gene Deletion and Chromosomal Tagging in Magnetospirillum gryphiswaldense. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 80(14), 4323-4330. doi:10.1128/AEM.00588-14.

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 Urheber:
Raschdorf, Oliver1, Autor           
Plitzko, Jürgen M.1, Autor           
Schüler, Dirk2, Autor
Müller, Frank D.2, Autor
Affiliations:
1Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565142              
2external, ou_persistent22              

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Schlagwörter: GRAM-NEGATIVE BACTERIA; MAGNETOSOME PROTEINS; MAGNETITE BIOMINERALIZATION; MAGNETOTACTIC BACTERIA; EXPRESSION; POLY(3-HYDROXYBUTYRATE); CONSTRUCTION; MUTAGENESIS; FILAMENTS; PARTICLES
 Zusammenfassung: Magnetotactic bacteria have emerged as excellent model systems to study bacterial cell biology, biomineralization, vesicle formation, and protein targeting because of their ability to synthesize single-domain magnetite crystals within unique organelles (magnetosomes). However, only few species are amenable to genetic manipulation, and the limited methods for site-specific mutagenesis are tedious and time-consuming. Here, we report the adaptation and application of a fast and convenient technique for markerless chromosomal manipulation of Magnetospirillum gryphiswaldense using a single antibiotic resistance cassette and galK-based counterselection for marker recycling. We demonstrate the potential of this technique by genomic excision of the phbCAB operon, encoding enzymes for polyhydroxyalkanoate (PHA) synthesis, followed by chromosomal fusion of magneto-some-associated proteins to fluorescent proteins. Because of the absence of interfering PHA particles, these engineered strains are particularly suitable for microscopic analyses of cell biology and magnetosome biosynthesis.

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Sprache(n): eng - English
 Datum: 2014-07
 Publikationsstatus: Erschienen
 Seiten: 8
 Ort, Verlag, Ausgabe: -
 Inhaltsverzeichnis: -
 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: ISI: 000338706800026
DOI: 10.1128/AEM.00588-14
 Art des Abschluß: -

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Titel: APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Genre der Quelle: Zeitschrift
 Urheber:
Affiliations:
Ort, Verlag, Ausgabe: 1752 N ST NW, WASHINGTON, DC 20036-2904 USA : AMER SOC MICROBIOLOGY
Seiten: - Band / Heft: 80 (14) Artikelnummer: - Start- / Endseite: 4323 - 4330 Identifikator: ISSN: 0099-2240