In vitro construction of different oligomeric forms of lambdadv DNA and studies of their transforming activities

Matsubara, Kouki; Takagi, Yasuyuki; Mukai, Tsunehiro

doi:10.1128/jvi.16.3.479-485.1975

Cited by 45 publications

(15 citation statements)

References 22 publications

(25 reference statements)

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“…Determination of the amount of covalently closed circular DNA (CCC DNA) plasmid per chromosome. The preparation of whole-cell lysates and the determination of plasmid copy number per chromosome were done as described by Matsubara et al (20). W3630 harboring Rms201 or Rms268 was grown to an optical density of 0.2 at 630 nm in 3 ml of L-broth at 37°C and labeled with 5 ,tCi of [3H]thymidine per ml or 2 iCi of ['4C]thymidine per ml for 90 min, respectively.…”

Section: Methodsmentioning

confidence: 99%

A mutant defective in partitioning of composite plasmid Rms201

Ike¹,

Hashimoto²,

Mitsuhashi³

1981

J Bacteriol

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derived from a Shigella strain (33) and conferred on its host resistance to tetracycline, chloramphenicol, streptomycin, sulfanilamide, ampicillin, spectinomycmn, and mercuric ions.Media and drugs. L-broth, bromothymol blue (BTB) lactose agar, Penassay broth, and Penassay broth agar were used. BTB agar consisted of nutrient broth (10 g of beef extract, 10 g of peptone, and 3 g of NaCl in 1 liter of distilled water) supplemented with 1% lactose, 0.008%' BTB, and 1.5% agar. Selective agar medium for conjugal transfer of the R plasmid from ML1410 R+ to W3630 was AG-agar. AG-agar medium consisted of medium A (7) with 1% glucose, 0.008'% BTB, and 1.5% agar. The selective concentration of drugs used for resistant bacteria were 12.5 pg of tetracycline, 12.5 pg of chloramphenicol, 6 pg of strepto-578 on August 1, 2020 by guest

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Section: Methodsmentioning

confidence: 99%

A mutant defective in partitioning of composite plasmid Rms201

Ike¹,

Hashimoto²,

Mitsuhashi³

1981

J Bacteriol

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“…and agar were used to grow and titrate bacteria and phage (10). PB medium was same as the polypeptone bonito extract broth medium, except that it contained 1 g instead of 10 g of bonito extract per liter (17). Minimal agar and miimal medium were supplemented with 20 jug of xanthine per ml, when necessary (10).…”

Section: Media Polvpeptone Bonito Extract Broth Mediun3mentioning

confidence: 99%

“…ColEl-cosA DNAs were accumulated by incubating cells in a minimal medium supplemented with 20 jig of xanthine per ml in the presence of 100 pg of chloramphenicol per ml (15). Extrachromosomal DNA was extracted and purified as described previously (17).…”

Section: Media Polvpeptone Bonito Extract Broth Mediun3mentioning

confidence: 99%

Lambda bacteriophage-mediated transduction of ColE1 deoxyribonucleic acid having a lambda bacteriophage-cohesive end site: selection of packageable-length deoxyribonucleic acid

Umene¹,

Shimada²,

Tsuzuki³

et al. 1979

J Bacteriol

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An in vitro recombinant ColEl-cosA deoxyribonucleic acid (DNA) molecule, pKY96, has 70% of the length of A phage DNA. The process of A phage-mediated transduction of pKY96 generated a small amount of transducing phage particles containing ColEl-cosA DNA molecules of 80 or 101% of the length of A phage DNA, in addition to those containing original pKY96 DNA molecules. The newly isolated larger plasmid DNAs were transduced 100 times more efficiently than pKY96 DNA. Their structures were compared with that of a prototype pKY96 DNA, and the mechanism of the formation of these molecules is discussed.GuaA gene transduction. A 0. 1-ml amount of KS1616 cell suspension in 0.01 M MgSOX and 0.1 ml of diluted phage lysates were mixed and incubated for :30 min at 30°C. The mixtures were plated on minimal agar supplemented with 20 tig of xanthine per ml, and the plates were examined after incub)ation for 2 days at 30°C (28).CsCl density gradient analysis. Phage Iysates on August 3, 2020 by guest

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“…The following method was used to determine the amount of plasmid covalently closed circular (CCC) DNA per chromosome. The preparation of whole-cell lysate and the determination of plasmid copy number per chromosome were done as described by Matsubara et al (18). Strain ML1410 cells harboring plasmids were grown to an optical density at 630 nm of 0.2 in 5 ml of L-broth at 3000 and were labeled with 5 jiCi of [3H]thymidine per ml for 90 min.…”

mentioning

confidence: 99%

Isolation and Characterization of a Composite Plasmid Rms201 Mutant Temperature Sensitive for Replication

et al. 1980

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A mutant temperature-sensitive for R-plasmid replication, Rms2O1tsl4, was isolated from composite plasmid Rms201 after mutagenesis of P1 transducing lysate with 100 mM hydroxylamine for 40 h at 370C. When Escherichia coli ML1410(Rms201ts14)+ was grown at temperatures between 40 and 420C in L broth, antibiotic-sensitive cells were segregated. When the incubation temperature of ML1410(Rms201ts14)+ in L-broth was shifted to 42 from 300C, the increase in the number of antibiotic-resistant cells ceased 90 min after the temperature shift. However, the total number of cells continuously increased, and only 3% of the cells retained the plasmid at 5 h after the temperature shift to 420C. At 300C the amounts of covalently closed circular deoxyribonucleic acid per chromosome of Rms2O1tsl4 and Rms201 were 3.8 and 6.3%, respectively. Incorporation of radioactive thymidine into the covalently closed circular deoxyribonucleic acid of Rms201tsl4 did not take place at 420C, whereas radioactive thymidine was incorporated into the covalently closed circular deoxyribonucleic acid of Rms201 at a rate of 4%/chromosome even at 420C. The synthesis of plasmid covalently closed circular deoxyribonucleic acid in a cell harboring Rms201tsl4 was almost completely blocked at 420C. These results indicated that the gene(s) responsible for plasmid deoxyribonucleic acid replication was affected in the mutant Rms2llts14. Temperature-sensitive miniplasmid pMSts214, which has a molecular weight of 5.3 x 106 and encodes ampicillin resistance, was isolated from Rms2O1tsl4. Similarly, miniplasnid pMS201, which encodes single ampicillin resistance, was isolated from its parent, Rms201, and its molecular weight was 4.7 x 106. These results indicate that the gene(s) causing temperature sensitivity for replication of Rms201 resides on the miniplasmid.

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In vitro construction of different oligomeric forms of lambdadv DNA and studies of their transforming activities

Cited by 45 publications

References 22 publications

A mutant defective in partitioning of composite plasmid Rms201

A mutant defective in partitioning of composite plasmid Rms201

Lambda bacteriophage-mediated transduction of ColE1 deoxyribonucleic acid having a lambda bacteriophage-cohesive end site: selection of packageable-length deoxyribonucleic acid

Isolation and Characterization of a Composite Plasmid Rms201 Mutant Temperature Sensitive for Replication

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