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      A New Method for Isolating Host-Independent Variants of Bdellovibrio bacteriovorus Using E. coli Auxotrophs

      research-article
      Author(s): , *
      Publication date (Electronic):
      Journal: The Open Microbiology Journal
      Publisher: Bentham Open
      Keywords: Bdellovibrio bacteriovorus, host-independent, isolation.

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          Abstract

          Bdellovibrios are Gram-negative bacteria that are characterized by predatory behavior. Although Bdellovibrios exhibit an obligatory parasitic life cycle, it is possible to isolate Bdellovibrio variants that no longer require host cells for their growth. In this study, a new method for isolating Bdellovibrio bacteriovorus host-independent (HI) variants was developed. Filtered B. bacteriovorus prey cells were cultured with E. coli diaminopimelic acid (DAP) auxotrophs as host cells. Thereafter, the lysate was plated on DAP minus media, allowing only HI colonies to develop. Using this method, we have isolated numerous HI variants and demonstrated that the emergence of HI variants may be occurring at a higher frequency than was previously suggested.

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          Genetic identification of a respiratory arsenate reductase.

          Chad W Saltikov, Dianne K Newman (2003)
          For more than a decade, it has been recognized that arsenate [H2AsO41-; As(V)] can be used by microorganisms as a terminal electron acceptor in anaerobic respiration. Given the toxicity of arsenic, the mechanistic basis of this process is intriguing, as is its evolutionary origin. Here we show that a two-gene cluster (arrAB; arsenate respiratory reduction) in the bacterium Shewanella sp. strain ANA-3 specifically confers respiratory As(V) reductase activity. Mutants with in-frame deletions of either arrA or arrB are incapable of growing on As(V), yet both are able to grow on a wide variety of other electron acceptors as efficiently as the wild-type. Complementation by the wild-type sequence rescues the mutants' ability to respire As(V). arrA is predicted to encode a 95.2-kDa protein with sequence motifs similar to the molybdenum containing enzymes of the dimethyl sulfoxide reductase family. arrB is predicted to encode a 25.7-kDa iron-sulfur protein. arrA and arrB comprise an operon that contains a twin arginine translocation (Tat) motif in ArrA (but not in ArrB) as well as a putative anaerobic transcription factor binding site upstream of arrA, suggesting that the respiratory As(V) reductase is exported to the periplasm via the Tat pathway and under anaerobic transcriptional control. These genes appear to define a new class of reductases that are specific for respiratory As(V) reduction.
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            Predation by Bdellovibrio bacteriovorus HD100 requires type IV pili.

            Adele Evans, Carey Lambert, R Sockett (2007)
            Early electron microscopy and more recent studies in our laboratory of Bdellovibrio bacteriovorus cells indicated the presence of narrow fibers at the nonflagellar pole of this unusual predatory bacterium. Analysis of the B. bacteriovorus HD100 genome showed a complete set of genes potentially encoding type IV pili and an incomplete gene set for Flp pili; therefore, the role of type IV pili in the predatory life cycle of B. bacteriovorus HD100 was investigated. Alignment of the predicted PilA protein with known type IV pilins showed the characteristic conserved N terminus common to type IVa pilins. The pilA gene, encoding the type IV pilus fiber protein, was insertionally inactivated in multiple Bdellovibrio replicate cultures, and the effect upon the expression of other pilus genes was monitored by reverse transcriptase PCR. Interruption of pilA in replicate isolates abolished Bdellovibrio predatory capability in liquid prey cultures and on immobilized yellow fluorescent protein-labeled prey, but the mutants could be cultured prey independently. Expression patterns of pil genes involved in the formation of type IV pili were profiled across the predatory life cycle from attack phase predatory Bdellovibrio throughout the intraperiplasmic bdelloplast stages to prey lysis and in prey-independent growth. Taken together, the data show that type IV pili play a critical role in Bdellovibrio predation.
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              Maintenance of broad-host-range incompatibility group P and group Q plasmids and transposition of Tn5 in Bartonella henselae following conjugal plasmid transfer from Escherichia coli.

              C Dehio, Allison M. Meyer (1996)
              The first demonstration of conjugal plasmid transfer from Escherichia coli to Bartonella henselae is reported. Transconjugants bearing plasmids of incompatibility groups P (IncP) and Q (IncQ), expressing various resistance markers, were generated. Tn5 transposons delivered on suicide plasmids by conjugation showed transpositional insertion into random chromosomal sites.
              Article 0 comments Cited 36 times – based on 0 reviews     Review now
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                Author and article information

                Journal
                Journal ID (nlm-ta): Open Microbiol J
                Journal ID (publisher-id): TOMICROJ
                Title: The Open Microbiology Journal
                Publisher: Bentham Open
                ISSN (Electronic): 1874-2858
                Publication date (Electronic): 2 June 2009
                Publication date Collection: 2009
                Volume: 3
                Pages: 87-91
                Affiliations
                []Department of Oral Biology, University of Medicine and Dentistry of New Jersey, Newark, NJ, 07101, USA
                Author notes
                [* ]Address correspondence to this author at the Department of Oral Biology, University of Medicine and Dentistry of New Jersey, Newark, NJ, 07101, USA; Tel: 973-972-7401; Fax: 973-972-0045; E-mail: Kadourde@ 123456umdnj.edu
                Article
                Publisher ID: TOMICROJ-3-87
                DOI: 10.2174/1874285800903000087
                PMC ID: 2705846
                PubMed ID: 19590595
                SO-VID: b3926798-ea13-4ae3-8cc6-cf22b551950c
                Copyright © © Dashiff and Kadouri; Licensee Bentham Open.
                License:

                This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

                History
                Date received : 5 May 2009
                Date revision received : 13 May 2009
                Date accepted : 15 May 2009
                Categories
                Subject: Article

                ScienceOpen disciplines: Microbiology & Virology
                Keywords: bdellovibrio bacteriovorus,host-independent,isolation.
                Data availability:
                ScienceOpen disciplines: Microbiology & Virology
                Keywords: bdellovibrio bacteriovorus, host-independent, isolation.

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