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      Effect of bacterial growth rate on bacteriophage population growth rate

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          Abstract

          It is important to understand how physiological state of the host influence propagation of bacteriophages (phages), due to the potential higher phage production needs in the future. In our study, we tried to elucidate the effect of bacterial growth rate on adsorption constant (δ), latent period (L), burst size (b), and bacteriophage population growth rate (λ). As a model system, a well‐studied phage T4 and Escherichia coli K‐12 as a host was used. Bacteria were grown in a continuous culture operating at dilution rates in the range between 0.06 and 0.98 hr −1. It was found that the burst size increases linearly from 8 PFU·cell −1 to 89 PFU·cell −1 with increase in bacteria growth rate. On the other hand, adsorption constant and latent period were both decreasing from 2.6∙10 ‐9 ml·min −1 and 80 min to reach limiting values of 0.5 × 10 ‐9 ml·min −1 and 27 min at higher growth rates, respectively. Both trends were mathematically described with Michaelis–Menten based type of equation and reasons for such form are discussed. By applying selected equations, a mathematical equation for prediction of bacteriophage population growth rate as a function of dilution rate was derived, reaching values around 8 hr −1 at highest dilution rate. Interestingly, almost identical description can be obtained using much simpler Monod type equation and possible reasons for this finding are discussed.

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          Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.

          A new method of total RNA isolation by a single extraction with an acid guanidinium thiocyanate-phenol-chloroform mixture is described. The method provides a pure preparation of undegraded RNA in high yield and can be completed within 4 h. It is particularly useful for processing large numbers of samples and for isolation of RNA from minute quantities of cells or tissue samples.
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            Molecular basis of bacterial outer membrane permeability revisited.

            Gram-negative bacteria characteristically are surrounded by an additional membrane layer, the outer membrane. Although outer membrane components often play important roles in the interaction of symbiotic or pathogenic bacteria with their host organisms, the major role of this membrane must usually be to serve as a permeability barrier to prevent the entry of noxious compounds and at the same time to allow the influx of nutrient molecules. This review summarizes the development in the field since our previous review (H. Nikaido and M. Vaara, Microbiol. Rev. 49:1-32, 1985) was published. With the discovery of protein channels, structural knowledge enables us to understand in molecular detail how porins, specific channels, TonB-linked receptors, and other proteins function. We are now beginning to see how the export of large proteins occurs across the outer membrane. With our knowledge of the lipopolysaccharide-phospholipid asymmetric bilayer of the outer membrane, we are finally beginning to understand how this bilayer can retard the entry of lipophilic compounds, owing to our increasing knowledge about the chemistry of lipopolysaccharide from diverse organisms and the way in which lipopolysaccharide structure is modified by environmental conditions.
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              Purification of RNA using TRIzol (TRI reagent).

              TRIzol solubilization and extraction is a relatively recently developed general method for deproteinizing RNA. This method is particularly advantageous in situations where cells or tissues are enriched for endogenous RNases or when separation of cytoplasmic RNA from nuclear RNA is impractical. TRIzol (or TRI Reagent) is a monophasic solution of phenol and guanidinium isothiocyanate that simultaneously solubilizes biological material and denatures protein. After solubilization, the addition of chloroform causes phase separation (much like extraction with phenol:chloroform:isoamyl alcohol), where protein is extracted to the organic phase, DNA resolves at the interface, and RNA remains in the aqueous phase. Therefore, RNA, DNA, and protein can be purified from a single sample (hence, the name TRIzol). TRIzol extraction is also an effective method for isolating small RNAs, such as microRNAs, piwi-associated RNAs, or endogeneous, small interfering RNAs. However, TRIzol is expensive and RNA pellets can be difficult to resuspend. Thus, the use of TRIzol is not recommend when regular phenol extraction is practical.
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                Author and article information

                Contributors
                ales.podgornik@cobik.si
                Journal
                Microbiologyopen
                Microbiologyopen
                10.1002/(ISSN)2045-8827
                MBO3
                MicrobiologyOpen
                John Wiley and Sons Inc. (Hoboken )
                2045-8827
                01 December 2017
                April 2018
                : 7
                : 2 ( doiID: 10.1002/mbo3.2018.7.issue-2 )
                : e00558
                Affiliations
                [ 1 ] Center of Excellence for Biosensors Instrumentation and Process Control ‐ COBIK Ajdovščina Slovenia
                [ 2 ] Faculty of Chemistry and Chemical Technology University of Ljubljana Ljubljana Slovenia
                Author notes
                [*] [* ] Correspondence

                Aleš Podgornik, Center of Excellence for Biosensors, Instrumentation and Process Control ‐ COBIK, Ajdovščina, Slovenia.

                Email: ales.podgornik@ 123456cobik.si

                Author information
                http://orcid.org/0000-0002-3080-737X
                Article
                MBO3558
                10.1002/mbo3.558
                5911998
                29195013
                f8ad4026-4789-441b-99f4-037d3f013700
                © 2017 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 24 July 2017
                : 12 October 2017
                : 13 October 2017
                Page count
                Figures: 3, Tables: 2, Pages: 10, Words: 8826
                Funding
                Funded by: Slovenian Research Agency (ARRS)
                Award ID: P3‐0387
                Award ID: 3030‐37543
                Award ID: L4‐5532
                Funded by: ARRS state budget
                Funded by: European Regional Development Fund
                Funded by: Slovenian Ministry of Education, Science and Sport
                Categories
                Original Research
                Original Research
                Custom metadata
                2.0
                mbo3558
                April 2018
                Converter:WILEY_ML3GV2_TO_NLMPMC version:version=5.3.4 mode:remove_FC converted:23.04.2018

                Microbiology & Virology
                bacterial growth rate,bacteriophage population growth rate,chemostat,e. coli k‐12,phage growth parameters,phage t4

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