Infection of Bacillus subtilis with phenol-extracted bacteriophages

Romig, W. R.

doi:10.1016/0042-6822(62)90226-x

Cited by 83 publications

(44 citation statements)

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“…Trypticase soy broth and agar (Baltimore Biological Laboratories, Baltimore, Maryland) were used for phage assays and propagation. TY broth [10] was modified for use in one-step growth procedures by reducing the concentration of sodium chloride to 0.5% and adding calcium chloride to a final concentration of 0.004 m. This broth was found to be the most satisfactory for one-step growth experiments since it yielded the least abortive infection. Agar for use with novobiocin-resistant bacterial cultures was prepared by adding novobiocin to standard trypticase soy agar to give a final concentration of 3 //g of novobiocin per ml of agar.…”

Section: Methodsmentioning

confidence: 99%

The Multiplication of Staphylococcal Phages in Nonpermissive Hosts

Wentworth

Romig

1968

Japanese Journal of Microbiology

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Section: Methodsmentioning

confidence: 99%

The Multiplication of Staphylococcal Phages in Nonpermissive Hosts

Wentworth

Romig

1968

Japanese Journal of Microbiology

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“…On utilise comme batteries hGtes, les souches B. subtilis 168-2 (ind-) (leu-) [16], isolee B partir de la souche B. subtilis 168 (ind-) [17] 8 M avec un tampon glycine-NaOH-NaC1 3 M, pH 9,2 (0,3 ml pour 2 ml de solution) et l'hydrolyse en mononucleotides 5'-OH est completee en incubant la solution 3 heures B 37", avec 100 pg/ml de phosphodiesterase de venin de serpent (Worthington) ; l'hydrolysat est plongb 5 min dans un bain-marie B 95" pour precipiter les proteines puis centrifuge; il La quantit6 de mononucleotides a BtB calculee en utilisant les coefficients d'extinction molaires suivants B pH4,25: AMP (E260= 15300)) GMP (~~~~= 1 2 8 0 0 ) , CMP ( E~~~ = 12400)) TMP ( E~~~ = 9 130), acide hydroxymethyluridylique ( E~~~ = 9 750).…”

Section: Souches Bacteriennesunclassified

Isolement et propriétés des chaînes du DNA de bactériophage 2 C

Truffaut

1970

European Journal of Biochemistry

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(Rep le 30 septembre/6 d6cembre 1969)Bacillus subtilis phage 2 C DNA is double-stranded, with a high molecular weight (about 100 million), a low T, (77.8"), and a high buoyant density (1.742 g/cm3) in a CsCl densitygradient.Denatured DNA centrifuged in a CsCl gradient reveals two bands (1.752 and 1.762 g/cm3) corresponding to the complementary strands.I n this communication, we report preparative separation of two strands from denatured DNA, either by chromatography on a methylated albumin-kieselguhr column, or by density-gradient centrifugation in the presence of poly G. The melting curves of native and denatured DNA, and of isolated fractions before and after mixing and annealing, are compared. As shown by spectrophotometry, and by buoyant density, a good renaturation was obtained by mixing and annealing the "light" and ('heavy') fractions ; each isolated fraction cannot be renatured alone.Poly G strongly binds to the heavy fraction, increasing its density in a CsCl gradient by about 48 mg/cm3; there is also some binding to the light fraction (density increase under the same conditions, 6 mg/cm3). The resulting density difference between the two fractions is more pronounced than the natural one: 52 mg/cm3 instead of 10 mg/cm3. Poly C only interacts with the light fraction and host ribosomal RNA with the heavy one.Native DNA, and both fractions isolated by centrifugation in the presence of poly G, were analysed for base composition using 32P-labelled DNA. Native DNA had a (G + C) content of 39.1As for the isolated fractions, there are more pyrimidines (cytosine, and hydroxymethyluracil replacing thymine) in the "heavier" fraction, which preferentially interacts with poly G.Both strands from infectious DNA were assayed for infectivity with negative results. The "renatured" DNA also gave negative results, probably owing to some natural or accidental breakage.Le

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“…These systems include @X 174-Escherichia coli bacteria [1,2,3,4], ADNA and helper phage-E. coli [5], ADNA-E. coli [ 6 ] , T,DNA and helper phage-E. coli [7], R 17 RNA-E . coli [8], SP8 DNA -BacilEus subtilis [9], and HP 1 DNA-Haemophilus influenzae [ …”

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Infectious Nucleic Acids of Escherichia coli Bacteriophages

Benzinger

Delius²,

Jaenisch

et al. 1967

European Journal of Biochemistry

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Various strains of Escherichia coli were grown to a titer of 5 x lo8 cells/ml in peptone media.Following osmotic shocks (peptone -+ water -+ 0.4 M sucrose) or lysozyme-EDTA treatment in hypertonic media, the spheroplasts were stored at 4" and then tested for competence with 20 minutes after the addition of a non-saturating level of SAPlabelled @X 174 DNA to such competent cell populations at 37", 10-20°/, of the label was adsorbed. Noncompetent cell populations adsorbed no more than 4O//, of the added label. Prolonging the DNA adsorption time did not give a higher efficiency of DNA infection, because the cells began losing competence through a temperature-dependent process.More than goo/, of the infectious @X 174 DNA centers remained sensitive to DNAase during the first 20 minutes in DNA adsorption medium. Plating @X 174 DNA and M 12 RNA-infected cells on eosin methylene blue agar during this period prevented the formation of phage, whereas the plaque-forming ability of the mature phage was not inhibited. These observations may suggest an unusual permeability of the competent cells.Inhibitors of phage RNA infection were removed by washing the competent cells. DNAase pretreatment of the spheroplasts increased their competence for infectious M 12 phage RNA.There has been a rapid succession of papers establishing the infectivity of a variety of phage DNA's and RNA for several hosts. These systems include @X 174-Escherichia coli bacteria [1,2,3,4] The procedures required for high infectious DNA yields vary with the type of host cell. In Bacillus subtilis and Haemophilus influenzae cultures many competent bacteria appear "spontaneously" during some stage of growth in special media. I n contrast, E. coli strains have a very low level of "spontaneous"Non-Standard Abbreviation. Plaque-forming units, p.f.u [7,12,13]. Osmotic shock of E . coli may also induce high levels of competence for infectious @X 174 DNA [I].In the present communication various methods of preparing competent E. coli bacteria are compared.Several improvements on earlier methods are also described; in particular, a preparation of frozen spheroplasts of constant competence is presented and methods for preparing spheroplasts of high

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Infection of Bacillus subtilis with phenol-extracted bacteriophages

Cited by 83 publications

References 8 publications

The Multiplication of Staphylococcal Phages in Nonpermissive Hosts

The Multiplication of Staphylococcal Phages in Nonpermissive Hosts

Isolement et propriétés des chaînes du DNA de bactériophage 2 C

Infectious Nucleic Acids of Escherichia coli Bacteriophages

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