In vivo isolation of a pPJ3a-λ cosmid mediated by the Tn2307 transposon

doi:10.1016/0147-619X(81)90034-2

Plasmid

Volume 6, Issue 3, November 1981, Pages 255-269

https://doi.org/10.1016/0147-619X(81)90034-2 Get rights and content

Abstract

From an E. coli cell harboring plasmid pPJ3b (= pPJ3a::Tn2301) and infected with phage λ, we have isolated two defective phages having inserted pPJ3a DNA and Tn2301 in their genomes. One of them has been extensively characterized: it behaves like a cosmid, i.e., upon injection into the cell, its DNA circularizes and replicates as a plasmid (pPJ10); it can be packaged again in λ heads, provided the presence of a phage helper. Furthermore, heteroduplex analysis has shown that in pPJ10, the transposon Tn2301 is inverted compared to its direction in pPJ3b. We give evidence suggesting that this type of inversion is in part mediated by Tn2301.

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Cited by (2)

TnA transposons can be introduced and maintained in Neisseria gonorrhoeae
1990, Research in Microbiology
In Neisseria gonorrhoeae, all penicillinase-specifying plasmids isolated so far share homology with each other and carry a 60 % deleted sequence of TnA. Plasmid pHD131, an element isolated from Haemophilus ducreyi and carrying an intact copy of the ampicillin resistance transposable element, was introduced from Escherichia coli into N. gonorrhoeae by both transformation and conjugative mobilization. Plasmids were recovered with no detectable deletion. After their transfer back into E. coli, transposition assays onto phage-γ DNA demonstrated that the TnA elements were still functional. Plasmid pHD131 could be stably maintained in N. gonorrhoeae with or without the presence of penicillin. These results support the hypothesis that the absence in N. gonorrhoeae of plasmids carrying entire and functional TnA transposons cannot be ascribed to incompatibility between the genetic element and the host, but rather to a barrier to introduction of foreign DNA into gonococcus.
Tous les plasmides codant pour des béta-lactamases et qui ont été isoleés jusqu'a présent chez les gonocoques sont homologues entre eux, et ils portent un même fragment correspondant à environ 40 % des transposons de type TnA. Le plasmide phD131 est un élément génétique isolé de Haemophilus ducreyi: il porte une copie intacte de TnA. A partir de Escherichia coli, nous avons introduit ce plasmide chez Neisseria gonorrhoeae par transformation ou par mobilisation conjugative, sans que l'on puisse détecter aucune délétion. Après son transfert en retour chez E. coli, la transposition de TnA sur le génome du phage 1 est toujours possible, ce qui démontre que la fonctionnalité du transposon est conservée. Nous avons pu montrer de plus que pHD131 peut être maintenu de façon stable chez le gonocoque. Ces résultats de transposons TnA intacts et fonctionnels ne peut pas être attribuée à une incompatibilité entre l'élément génétique et l'hôte mais plutôt à une barrière empêchant l'introduction d'ADN étranger chez le gonocoque.
An F-derived conjugative cosmid: Analysis of tra polypeptides in cosmid-infected cells
1986, Plasmid
The genes involved in the conjugational transfer of F plasmid DNA are organized into three closely linked operons spanning an overall length of approximately 33 kilobase pairs of F. The entire transfer (tra) region comprising all three operons has been cloned into the cosmid vector pHC79 by in vitro recombination and packaging techniques. The transfer-proficient chimeric cosmid pRS2405 was packaged into λ capsids, and uv-irradiated E. coli cells were infected with these DNA-filled particles. A number of polypeptides programmed by the infecting DNA were identified as tra-specified products; a traJ90 mutation on pRS2405 resulted in the significant reduction of synthesis of all detectable pRS2405-specified tra polypeptides, with the exception of TraTp.

¹: Present address and reprints: Istituto Cantonale Batteriologico, Via Ospedale 6, 6904 Lugano, Switzerland.

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Plasmid

Abstract

References (28)

Construction and characterization of new cloning vehicles. I. Ampicillin-resistant derivatives of the plasmid pMB9

Gene

Physical mapping of coliphage λatt

Virology

DNA sequence analysis of the transposon Tn3: three genes and three sites involved in transposition of Tn3

Cell

DNA as a substrate for packaging into bacteriophage lambda, in vitro

J. Mol. Biol

Site-specific DNA deletions occurring adjacent to the termini of a transposable ampicillin resistance element. (Tn3)

J. Mol. Biol

Phage lambda mediated transduction of resistance plasmids promoted by Ap^R transposons

Gene

Detection of specific sequences among DNA fragments separated by gel electrophoresis

J. Mol. Biol

Processing of bacteriophage lambda DNA during its assembly into heads

J. Mol. Biol

Fragments produced by cleavage of λ deoxyribonucleic acid with the Haemophilus parainfluenzae restriction enzyme HpaII

Biochemistry

Dissection of the transposition process: A transposon-encoded site-specific recombination system

Mol. Gen. Genet

Transposition of R-factor genes to bacteriophage λ

Involvement of IS1 in the dissociation of the r-determinant and RTF components of the plasmid R100.1

Mol. Gen. Genet

Transposition protein of Tn3: identification and characterization of an essential repressor-controlled gene product

Nature (London)

Super-coiled circular DNA-protein complex in E. coli: Purification and induced conversion to an open circular form

Cited by (2)

TnA transposons can be introduced and maintained in Neisseria gonorrhoeae

An F-derived conjugative cosmid: Analysis of tra polypeptides in cosmid-infected cells

Regular articleIn vivo isolation of a pPJ3a-λ cosmid mediated by the Tn2307 transposon

Abstract

Gene

Virology

Cell

J. Mol. Biol

J. Mol. Biol

Gene

J. Mol. Biol

J. Mol. Biol

Fragments produced by cleavage of λ deoxyribonucleic acid with the Haemophilus parainfluenzae restriction enzyme HpaII

Biochemistry

Dissection of the transposition process: A transposon-encoded site-specific recombination system

Mol. Gen. Genet

Transposition of R-factor genes to bacteriophage λ

Involvement of IS1 in the dissociation of the r-determinant and RTF components of the plasmid R100.1

Mol. Gen. Genet

Transposition protein of Tn3: identification and characterization of an essential repressor-controlled gene product

Nature (London)

Super-coiled circular DNA-protein complex in E. coli: Purification and induced conversion to an open circular form

Regular article
In vivo isolation of a pPJ3a-λ cosmid mediated by the Tn2307 transposon