title: Assessment of the mobilizable vector plasmids pSUP202 and pSUP404.2 as genetic tools for the predatory bacterium Bdellovibrio bacteriovorus.
abstract: Bdellovibrio and like organisms (BALOs) form the group of predatory bacteria which require Gram-negative bacteria as prey. Genetic studies with Bdellovibrio bacteriovorus can be performed with vectors which are introduced into the predator via conjugation. The usefulness of the two vectors pSUP202 and pSUP404.2 as genetic tools were assessed. Both vectors were transferable into B. bacteriovorus by conjugative matings with an Escherichia coli K12 strain as donor. The transfer frequency was higher for vector pSUP404.2 (approx. 10⁻¹-10⁻⁴) as for pSUP202 (approx. 10⁻⁵-10⁻⁶). Vector pSUP202 with a pMB1 origin is unstable in the predatory bacterium, whereas pSUP404.2 is stably maintained in the absence of selective antibiotics. pSUP404.2 harbors two plasmid replicons, the p15A ori and the RSF1010 replication region The copy number of pSUP404.2 was determined by quantitative PCR in B. bacteriovorus and averages seven copies per genome. pSUP404.2 harbors two resistance genes (chloramphenicol and kanamycin) which can be used for cloning either by selection for transconjugants or by insertional inactivation.
authors.affiliation: Department of Biological Safety, Bundesinstitut für Risikobewertung (BfR), Federal Institute for Risk Assessment, Diedersdorfer Weg 1, 12277, Berlin, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Roschanski
authors.forename: Eckhard
authors.initials: E
authors.lastname: Strauch
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001501:Bdellovibrio / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D003227:Conjugation, Genetic; D048168:Escherichia coli K12 / Q000235:genetics; D022761:Gene Transfer, Horizontal; D005822:Genetic Vectors; D042822:Genomic Instability; D008969:Molecular Sequence Data; D010957:Plasmids; D012641:Selection, Genetic; D017422:Sequence Analysis, DNA
chemical_list: D000900:Anti-Bacterial Agents
references.citation: J Bacteriol. 1992 Oct;174(19):6011-7
references.pmid: 1400153
references.citation: J Biotechnol. 2000 Apr 14;79(1):63-72
references.pmid: 10817342
references.citation: J Bacteriol. 2010 Mar;192(5):1299-311
references.pmid: 20023029
references.citation: Appl Environ Microbiol. 2002 Dec;68(12):5834-42
references.pmid: 12450802
references.citation: Cell. 1983 Jan;32(1):119-29
references.pmid: 6186390
references.citation: Int J Syst Evol Microbiol. 2004 Sep;54(Pt 5):1439-1452
references.pmid: 15388693
references.citation: Science. 2004 Jan 30;303(5658):689-92
references.pmid: 14752164
references.citation: J Bacteriol. 1985 Jul;163(1):324-30
references.pmid: 3891734
references.citation: Gene. 1982 Jan;17(1):79-89
references.pmid: 6281139
references.citation: Syst Appl Microbiol. 2001 Nov;24(3):385-94
references.pmid: 11822674
references.citation: J Bacteriol. 1969 Feb;97(2):912-23
references.pmid: 4886299
references.citation: Gene. 1991 Jan 2;97(1):39-47
references.pmid: 1847347
references.citation: Microbiol Rev. 1984 Mar;48(1):1-23
references.pmid: 6201704
references.citation: Microbiology (Reading). 2004 Mar;150(Pt 3):649-656
references.pmid: 14993314
references.citation: Appl Environ Microbiol. 2008 Sep;74(17):5436-43
references.pmid: 18621871
references.citation: Appl Environ Microbiol. 2007 Aug;73(15):4717-24
references.pmid: 17557848
references.citation: Future Microbiol. 2007 Feb;2(1):63-73
references.pmid: 17661676