Biphasic System for Growing Bacteria in Concentrated Culture

Tyrrell, Elizabeth; Donald, Russell E. Mac; Gerhardt, Philipp

doi:10.1128/jb.75.1.1-4.1958

Cited by 45 publications

(16 citation statements)

References 4 publications

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“…Both strains have an LD SH in mice of V10 microorganisms when administered intraperitoneally. Cultures were grown to post-exponential phase in Luria-Bertani broth (Difco) at 37³C under anaerobic, nonagitated conditions, (5% CO P , 10% H P , 85% N P ) using a bi-phasic batch culture method [11] 2.3. Envelope protein preparations from S. typhimurium Bacterial cultures were harvested, concentrated, washed twice, and resuspended in ice-cold sonication bu¡er consisting of 10 mM Tris-HCl, pH 7.4, 5 mM MgCl P , 1.0 mM dithiothreitol (DTT), 50 Wg ml 3I each of ribonuclease A and deoxyribonuclease I (Worthington Biochemical Corp., Freehold, NJ), 10 Wg ml 3I leupeptin, 10 Wg ml 3I pepstatin, 1.0 mM phenylmethylsulfonyl £uoride, and 1.0% aprotinin (Calbiochem, San Diego, CA).…”

Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

Macrophages recognize and adhere to an OmpD-like protein of Salmonella typhimurium

Negm

1998

FEMS Immunology and Medical Microbiology

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Murine peritoneal macrophages bind to Salmonella typhimurium in vitro in the absence of exogenous opsonins. We have identified an outer membrane protein of S. typhimurium that mediates this adhesion. Biotin-labeled macrophages were used to probe electroblotted envelope proteins of S. typhimurium that had been previously resolved by polyacrylamide electrophoresis under denaturing and reducing conditions. Macrophages bound to an outer membrane protein with an apparent molecular mass of 44 kDa. The protein was purified to homogeneity and free of detectable lipopolysaccharide. Limited microsequencing of this protein resulted in a 15-amino acid query sequence of A-E-V-Y-N-K-D-G-N-K-L-D-L-Y-G, which shares complete identity with a 15-mer of both the OmpD of S. typhimurium SH 7454 and the OmpC polypeptide of Escherichia coli K-12. Picomolar concentrations of this purified protein significantly inhibited the subsequent adherence of 35S-labeled S. typhimurium to macrophages in monolayers. We propose that this 44-kDa protein is involved in the recognition of S. typhimurium by macrophage during the initial stages of infection.

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Section: Bacterial Strains and Growth Conditionsmentioning

confidence: 99%

Macrophages recognize and adhere to an OmpD-like protein of Salmonella typhimurium

Negm

1998

FEMS Immunology and Medical Microbiology

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“…For batch cultivation, broth cultures were shaken for 16 h with continuous gassing. Simon (17) briefly reported successful cultivation of a laboratory strain of Campylobacter fetus in a diphasic medium, a culture system shown by Tyrrell et al (23) to be advantageous for the growth of other genera.…”

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confidence: 99%

Biphasic culture system for rapid Campylobacter cultivation

Rollins¹,

Coolbaugh²,

Walker³

et al. 1983

Appl Environ Microbiol

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We developed a biphasic culture system consisting of 4 ml of brucella agar (BA) and 6 ml of brucella broth (BB) in 25-cm2 tissue culture flasks, which were incubated in air (BB/BAa) or in a gas mixture of 5% 02, 10% CO2, and 85% N2 (BB/BAg). These media were also used with a supplement consisting of ferrous sulfate, sodium metabisulfite, and sodium pyruvate and incubated as above (FB/ FAa and FB/FAg, respectively). Highly satisfactory growth of Campylobacter jejuni 301 was obtained with all medium-gas phase combinations provided that the number of viable cells in the inoculum was large (-106/ml). The use of FB/FAa permitted the inoculum to be reduced to 100 cells per ml. With an adjusted gas

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“…Continuous cultures are another alternative, but considerable time is required to establish proper conditions. In principle it should also be possible to grow high-density cultures in larger volumes than reported by Gerhardt's group (4,15,17). Alcaligenes eutrophus was eminently suited for these studies because it grows autotrophically in a defined inorganic medium with no significant production of metabolic waste products.…”

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confidence: 99%

Quantitative requirements for exponential growth of Alcaligenes eutrophus

Repaske¹,

Repaske²

1976

Appl Environ Microbiol

109

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Quantitative nutrient requirements for unrestricted autotrophic growth of Alcaligenes eutrophus were determined. Minimum saturating concentrations of Mg2+, S042, P043, Fe3+, and Na2+ for an optical density increase of 2 were 10-M, 8 x 10-5 M, 5 x 10-4 to 6 x 10-4 M, 10-5 M, and 10-7 to 2 x 10-7 M, respectively. Trace metal requirements for cobalt, chromium, and copper were also demonstrated, but minimum concentrations could not be determined because other reagents contributed a high background of these metals. Under certain conditions an apparent response to zinc was observed, although other experiments suggest the zinc salt contained another metal that was required for growth. Poly-,8-hydroxybutyrate biosynthesis was shown to be initiated by a magnesium or sulfate deficiency as well as by a nitrogen or phosphate deficiency.

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Biphasic System for Growing Bacteria in Concentrated Culture

Cited by 45 publications

References 4 publications

Macrophages recognize and adhere to an OmpD-like protein of Salmonella typhimurium

Macrophages recognize and adhere to an OmpD-like protein of Salmonella typhimurium

Biphasic culture system for rapid Campylobacter cultivation

Quantitative requirements for exponential growth of Alcaligenes eutrophus

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