Isolation of covalently closed circular DNA of high molecular weight from bacteria

Currier, Thomas C.; Nester, Eugene W.

doi:10.1016/0003-2697(76)90338-9

Cited by 405 publications

(166 citation statements)

References 11 publications

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“…To identify fragments homologous to IS66, hybridizations with cloned EcoRI fragments of pTiB6806 were also carried out. The probes weakly hybridized with cloned EcoRI fragments 2 (12 kbp, lane a) and 24 -k) showed, in each case, intense hybridization to fragment 11 (5.5-kbp) and weak hybridization to both fragment 2 (16-kbp) and fragment 3 (13.5-kbp). Therefore, the octopine Ti plasmids contain at least three sequences homologous to IS66.…”

Section: Resultsmentioning

confidence: 94%

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Nucleotide sequence of the insertion sequence found in the T-DNA region of mutant Ti plasmid pTiA66 and distribution of its homologues in octopine Ti plasmid.

Machida¹,

Sakurai²,

Kiyokawa³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

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The octopine tumor-inducing (Ti) plasmid pTiA66 has an insertion mutation in its T region (the DNA region incorporated into the plant genome) that results in the slow growth of crown gall tumors. These tumors exhibit hormonal autonomy different from that of the crown gall tumors caused by wild-type Ti plasmids. In the present study, the nucleotide sequences of both the DNA segment inserted into pTiA66 and its target site have been determined. The inserted segment is 2548 base pairs long and has 20-base-pair terminal inverted repeats. An 8-base-pair sequence at the target site is duplicated at both integration junctions. These structural features of the insert suggest that it is a bacterial insertion sequence (IS) element, which we have named IS66. Blot-hybridization analyses using IS66 probes revealed that genomes of octopine Ti plasmids contain at least three sequences homologous to IS66: two homologues are located in the virulence region and one is located between the left-hand (TL-DNA) and right-hand (TR-DNA) portions of T-DNA. The chromosome of Agrobacterium tumefaciens A66 also contains two sequences highly homologous to IS66. These results suggest that the mutant pTiA66 plasmid was generated by translocation of one of the sequences showing homology with IS66 into the T region. The fact that a sequence homologous to IS66 is present between TL-DNA and TR-DNA also suggests that the octopine T region was split into two portions, TL-DNA and TR-DNA, by translocation of IS66 or its relatives. Thus, IS66 may cause genetic and structural variations of the T region and the vir region of the octopine Ti plasmids.Tumor-inducing (Ti) plasmids harbored by oncogenic strains of Agrobacterium tumefaciens cause formation of tumors on dicotyledonous plants, called crown galls (1). Cells isolated from the crown gall tumors are characterized by their unlimited proliferation in medium lacking phytohormones, such as cytokinins and auxins, that are required for the culture of normal plant cells (2). It has been shown that a specific DNA region of the Ti plasmid, called the T-DNA region (Fig. 1), causes tumor formation when transferred into plant chromosomes and that the T-DNA persists in the chromosomes of the tumor cells (3-6). The virulence (vir) region of the Ti plasmid (Fig. 1) is located outside the T region and is also required for tumorigenesis by the Ti plasmid. However, this region is probably not stably maintained in the tumor cells (7-10).The Ti plasmids are classified on the basis of the novel amino acid metabolites, such as octopine or nopaline, whose syntheses are directed by T-DNA genes in the tumor cells (11-15). The octopine T region is composed of twQ DNA segments near each other in the Ti plasmid (16) (Fig. 1), whereas the nopaline T region consists of one contiguous DNA segment (17, 18). The chromosomes of the tumor cells caused by the octopine strain contain either one or two portions of the T region (16). All tumor cells that have been analyzed contained the left portion of T-DNA (TL-DNA) but not all...

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Section: Resultsmentioning

confidence: 94%

“…Escherichia coli C600 (rk-mk-thi thr leuB typB) used for transformation was described by Nagahari et al (22) (24), except that Pronase and phenol treatments were carried out in half the volume of the original bacterial culture. E. coli plasmid DNA was prepared by the alkaline method (25).…”

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confidence: 99%

Nucleotide sequence of the insertion sequence found in the T-DNA region of mutant Ti plasmid pTiA66 and distribution of its homologues in octopine Ti plasmid.

Machida¹,

Sakurai²,

Kiyokawa³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

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“…Synthesis of double-stranded cDNA (Maniatis et al, 1976), transformation of Escherichia coli x 1776 by pBR322-cDNA hybrid (Enea et al, 1975) and detection of the specific recombinant DNA by in situ colony hybridization (Grunstein and Hogness, 1975) were performed essentially according to the published procedures. Preparation of plasmid DNA was according to the procedure of Currier and Nester (1976) with some modifications.…”

Section: Introductionmentioning

confidence: 99%

Construction and identification of a bacterial plasmid containing the human fibroblast interferon gene sequence.

Taniguchi

Sakai

Fujii‐Kuriyama

et al. 1979

Proceedings of the Japan Academy. Ser. B: Physical and Biologic

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“…13 by the Currier & Nester (1976) procedure, as modified above, was directly applied to Spinco SW41 log-linear alkaline sucrose gradients and centrifuged as above. During fractionation the A254 was monitored on an ISCO model UA-4 absorbance monitor (Instrumentation Specialties Co.) and then 3H was measured.…”

Section: Introductionmentioning

confidence: 99%

Indigenous Plasmids from Phytopathogenic Corynebacterium Species

Gross

Vidaver

Keralis

1979

Journal of General Microbiology

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Efficient and reproducible cell lysis and the isolation of plasmid DNA from nine phytopathogenic Corynebacterium species is described. One to three plasmids, ranging from 23 to 77 megadaltons, were recovered from each strain, as detected by comparative agarose gel electrophoresis. Corynebacterium michiganense showed considerable plasmid diversity. Other species, such as C. nebraskense and C. insidiosum, did not. Bacteriocin production, colony morphology, pigmentation and virulence were not correlated with the presence of plasmids. Sedimentation through neutral and alkaline sucrose gradients and electrophoretic separation on agarose gels gave similar values for plasmid molecular weights. Log-linear alkaline sucrose gradients proved useful for molecular weight determinations and also for the isolation of purified plasmids on a preparative scale.

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Isolation of covalently closed circular DNA of high molecular weight from bacteria

Cited by 405 publications

References 11 publications

Nucleotide sequence of the insertion sequence found in the T-DNA region of mutant Ti plasmid pTiA66 and distribution of its homologues in octopine Ti plasmid.

Nucleotide sequence of the insertion sequence found in the T-DNA region of mutant Ti plasmid pTiA66 and distribution of its homologues in octopine Ti plasmid.

Construction and identification of a bacterial plasmid containing the human fibroblast interferon gene sequence.

Indigenous Plasmids from Phytopathogenic Corynebacterium Species

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