Transitory recombination between plasmid pHV33 and phage M13.

Dagert, M.; Ehrlich, S. Dusko

doi:10.1002/j.1460-2075.1983.tb01711.x

Cited by 10 publications

(13 citation statements)

References 26 publications

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“…The terminal-labeling method was used for DNA sequencing (32). Transformation of B. subtilis and E. coli competent cells and plasmid transduction have been described (28,33,34).…”

Section: Methodsmentioning

confidence: 99%

Illegitimate recombination at the replication origin of bacteriophage M13.

Michel¹,

Ehrlich²

1986

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

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Hybrids composed of phage M13 and plasmid pHV33 were used to study the formation of deletions in Escherchia coli. Eighty to ninety percent of the deletion endpoints were at the position of the nick introduced into the M13 replication origin by the phage gene II protein. This suggests the existence of a novel mechanism of illegitimate recombination.Recombination between sequences with little or no homology was termed "illegitimate" by Franklin (1) ref. 14), and AB1157 (thrAl leu-6 thi-) proA2 his4 argE3 lacYl galK2 ara-14 xyl-S mtl-i tsx-33 rpl-31 supE44; J.Clark, University of California, Berkeley). The Rec-strains were all isogenic to AB1157. They were rendered hsdR by conjugation with GY2098 (HfrH) after cotransfer of hsdR and thr'. recA strains JC5495 and JC5547 were rendered Rec' prior to conjugation by introduction of a RecAI X phage. This phage was eliminated from the exconjugants by segregation. All the Rec-strains harbored F'lacIqZ M15 pro' plasmid (15). The Rec-phenotype was routinely tested by sensitivity to ultraviolet light. The Rep-strain D162 was rendered hsdR by conjugation with a leu+ revertant of GY2098 after cotransfer of hsdR and leu+ markers. It harbored pOXKm plasmid (M. Chandler, Toulouse University).The plasmid pHV33 is composed of pBR322 and pC194 (16-18). The plasmids pHV672 (19) and pHV673 were constructed by joining EcoRI-cleaved M13 mp2 phage (20) to pHV33; they were isolated in B. subtilis. pHV698 was constructed by replacing the small pBR322 Cla I-EcoRI fragment with the 540-base-pair (bp) M13 mp2 Cla I-EcoRI fragment that carries the phage replication origin. pHV702 was obtained by inserting pC194 into the HindIII site of pHV698. R199 and R229 are two fl derivatives (21,22) (28,29) and was purified by chromatography on hydroxyapatite columns (30). For analytical purposes, plasmid DNA was extracted from 1.5-ml overnight cultures (31). The terminal-labeling method was used for DNA sequencing (32). Transformation of B. subtilis and E. coli competent cells and plasmid transduction have been described (28,33,34). RESULTSIsolation of Deletion Plasmids. Plasmid pHV33, composed ofthe E. coli plasmid pBR322 and the Staphylococcus aureus plasmid pC194, is viable in E. coli (18). We have joined the Abbreviations: bp, base pair(s); kb, kilobase(s). 3386The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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“…The terminal-labeling method was used for DNA sequencing (32). Transformation of B. subtilis and E. coli competent cells and plasmid transduction have been described (28,33,34).…”

Section: Methodsmentioning

confidence: 99%

Illegitimate recombination at the replication origin of bacteriophage M13.

Michel¹,

Ehrlich²

1986

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

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“…of the chimeras. These chimeras are supposed to be composed of one phage and one plasmid genome, as was previously shown for chimeras between M13 and pHV33 (Dagert and Ehrlich, 1983). Broken line is interpolated between the mobilities of less intense bands, solid line between those of more intense bands and that of the faster pBR322 band.…”

Section: Resultsmentioning

confidence: 99%

“…Plasmids transduced by M13 undergo a process which we call transitory recombination, and within which we distinguish two phases (Dagert and Ehrlich, 1983). During the first phase the plasmid and phage genomes combine, and form a chimera, during the second the chimera decombines to regenerate the parental genomes.…”

Section: Discussionmentioning

confidence: 99%

“…It occurred at a rather low frequency (10-11 transductants/phage) and yielded recombinants (Ravetch et al, 1979) between the two genomes by a process akin to the formation of co-integrates during transposition (Fischoff et al, 1980;Kleckner, 1981). We have recently described transduction of plasmid pHV33 (Primrose and Ehrlich, 1981) by M13, which occurred at much higher frequency (10-4 transductants/phage), but did not give rise to stable recombinants (Dagert and Ehrlich, 1983). This observation led us to postulate the existence of a previously undetected process, which we named transitory recombination, and in which we distinguished two phases.…”

Section: Introductionmentioning

confidence: 99%

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Replication functions of pC194 are necessary for efficient plasmid transduction by M13 phage.

Dagert¹,

Jones²,

Goze³

et al. 1984

The EMBO Journal

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Escherichia coli plasmids pBR313 and pBR322 were transduced by phage M13 with low efficiency (10(‐8) transductants/phage). Hybrid plasmids pHV12 or pHV33, composed of Staphylococcus aureus plasmid pC194 and pBR313 or pBR322, respectively, were transduced much more efficiently (10(‐4) transductants/phage). Inactivation of either of the two zones necessary for pC194 replication, one coding for a protein, the other not, reduced the transforming efficiency of hybrids to the level of pBR322. Activity of the pC194 replication region was not necessary for the formation of chimeras between M13 and the transduced plasmid in the donor cells, but rather for the establishment of the plasmid in the recipient cells.

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“…ApRKmR colonies were found with a frequency of about 1 x 10 ·· 7 • In control experiments using BHB3169[pAN26] in combination with the human genomic library ApR Km R colonies appeared with about the same frequency (Table 1). We do not know whether this is due to a type of transient recombination as described for M 13 phages (Dagert and Ehrlich, 1983) or to the presence of sequences homologaus to pAN26 in the DNA library. But in spite of this nonspecific background, truc recombinants should be highly enriched in the doubly resistant colonies.…”

Section: Results and Discussjon (A) Recombinant Screening Of Cosmid Lmentioning

confidence: 99%

Isolation of a functional human interleukin 2 gene from a cosmid library by recombination in vivo

Lindenmaier

Dittmar

Hauser

et al. 1985

Gene

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SUMMARYA method has been developed that allows the isolation of genomic clones from a cosmid library by homologaus recombination in vivo. This method was used to isolate a human genomic interleukin 2 (IL2) gene. The genomic cosmid library was packaged in vivo into A. phage particles. A recombination-proficient host strain carrying IL2 cDNA sequences in a non-homologaus plasmid vector was infected by the packaged cosmid library. After in vivo packaging and reinfection, recombinants carrying the antibiotic resistance genes of both vectors were selected. From a recombinant cosmid clone the chromosomal IL2 genewas restored. After DNA mediated gene transfer into mouse Ltk-cells human IL2 was expressed constitutively. JNTRODUCTION

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Transitory recombination between plasmid pHV33 and phage M13.

Cited by 10 publications

References 26 publications

Illegitimate recombination at the replication origin of bacteriophage M13.

Illegitimate recombination at the replication origin of bacteriophage M13.

Replication functions of pC194 are necessary for efficient plasmid transduction by M13 phage.

Isolation of a functional human interleukin 2 gene from a cosmid library by recombination in vivo

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