Site-specific deletions of chromosomally located DNA segments with the multimer resolution system of broad-host-range plasmid RP4

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Site-specific deletions of chromosomally located DNA segments with the multimer resolution system of broad-host-range plasmid RP4. / Kristensen, C S; Eberl, Leo; Sanchez-Romero, J M; Givskov, M; Molin, Søren; De Lorenzo, V.

In: Journal of Bacteriology, Vol. 177, No. 1, 1995, p. 52-8.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Kristensen, CS, Eberl, L, Sanchez-Romero, JM, Givskov, M, Molin, S & De Lorenzo, V 1995, 'Site-specific deletions of chromosomally located DNA segments with the multimer resolution system of broad-host-range plasmid RP4', Journal of Bacteriology, vol. 177, no. 1, pp. 52-8.

APA

Kristensen, C. S., Eberl, L., Sanchez-Romero, J. M., Givskov, M., Molin, S., & De Lorenzo, V. (1995). Site-specific deletions of chromosomally located DNA segments with the multimer resolution system of broad-host-range plasmid RP4. Journal of Bacteriology, 177(1), 52-8.

Vancouver

Kristensen CS, Eberl L, Sanchez-Romero JM, Givskov M, Molin S, De Lorenzo V. Site-specific deletions of chromosomally located DNA segments with the multimer resolution system of broad-host-range plasmid RP4. Journal of Bacteriology. 1995;177(1):52-8.

Author

Kristensen, C S ; Eberl, Leo ; Sanchez-Romero, J M ; Givskov, M ; Molin, Søren ; De Lorenzo, V. / Site-specific deletions of chromosomally located DNA segments with the multimer resolution system of broad-host-range plasmid RP4. In: Journal of Bacteriology. 1995 ; Vol. 177, No. 1. pp. 52-8.

Bibtex

@article{a5db4dec1d30490ba868377f788f5cc7,
title = "Site-specific deletions of chromosomally located DNA segments with the multimer resolution system of broad-host-range plasmid RP4",
abstract = "The multimer resolution system (mrs) of the broad-host-range plasmid RP4 has been exploited to develop a general method that permits the precise excision of chromosomal segments in a variety of gram-negative bacteria. The procedure is based on the site-specific recombination between two directly repeated 140-bp resolution (res) sequences of RP4 effected by the plasmid-borne resolvase encoded by the parA gene. The efficiency and accuracy of the mrs system to delete portions of chromosomal DNA flanked by res sites was monitored with hybrid mini-Tn5 transposons in which various colored (beta-galactosidase and catechol 2,3 dioxygenase) or luminescent (Vibrio harveyi luciferase) phenotypic markers associated to res sequences were inserted in the chromosome of the target bacteria and exposed in vivo to the product of the parA gene. The high frequencies of marker excision obtained with different configurations of the parA expression system suggested that just a few molecules of the resolvase are required to achieve the site-specific recombination event. Transient expression of parA from a plasmid unable to replicate in the target bacterium was instrumental to effect differential deletions within complex hybrid transposons inserted in the chromosome of Pseudomonas putida. This strategy permits the stable inheritance of heterologous DNA segments virtually devoid of the sequences used initially to select their insertion.",
author = "Kristensen, {C S} and Leo Eberl and Sanchez-Romero, {J M} and M Givskov and S{\o}ren Molin and {De Lorenzo}, V",
year = "1995",
language = "English",
volume = "177",
pages = "52--8",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "1",

}

RIS

TY - JOUR

T1 - Site-specific deletions of chromosomally located DNA segments with the multimer resolution system of broad-host-range plasmid RP4

AU - Kristensen, C S

AU - Eberl, Leo

AU - Sanchez-Romero, J M

AU - Givskov, M

AU - Molin, Søren

AU - De Lorenzo, V

PY - 1995

Y1 - 1995

N2 - The multimer resolution system (mrs) of the broad-host-range plasmid RP4 has been exploited to develop a general method that permits the precise excision of chromosomal segments in a variety of gram-negative bacteria. The procedure is based on the site-specific recombination between two directly repeated 140-bp resolution (res) sequences of RP4 effected by the plasmid-borne resolvase encoded by the parA gene. The efficiency and accuracy of the mrs system to delete portions of chromosomal DNA flanked by res sites was monitored with hybrid mini-Tn5 transposons in which various colored (beta-galactosidase and catechol 2,3 dioxygenase) or luminescent (Vibrio harveyi luciferase) phenotypic markers associated to res sequences were inserted in the chromosome of the target bacteria and exposed in vivo to the product of the parA gene. The high frequencies of marker excision obtained with different configurations of the parA expression system suggested that just a few molecules of the resolvase are required to achieve the site-specific recombination event. Transient expression of parA from a plasmid unable to replicate in the target bacterium was instrumental to effect differential deletions within complex hybrid transposons inserted in the chromosome of Pseudomonas putida. This strategy permits the stable inheritance of heterologous DNA segments virtually devoid of the sequences used initially to select their insertion.

AB - The multimer resolution system (mrs) of the broad-host-range plasmid RP4 has been exploited to develop a general method that permits the precise excision of chromosomal segments in a variety of gram-negative bacteria. The procedure is based on the site-specific recombination between two directly repeated 140-bp resolution (res) sequences of RP4 effected by the plasmid-borne resolvase encoded by the parA gene. The efficiency and accuracy of the mrs system to delete portions of chromosomal DNA flanked by res sites was monitored with hybrid mini-Tn5 transposons in which various colored (beta-galactosidase and catechol 2,3 dioxygenase) or luminescent (Vibrio harveyi luciferase) phenotypic markers associated to res sequences were inserted in the chromosome of the target bacteria and exposed in vivo to the product of the parA gene. The high frequencies of marker excision obtained with different configurations of the parA expression system suggested that just a few molecules of the resolvase are required to achieve the site-specific recombination event. Transient expression of parA from a plasmid unable to replicate in the target bacterium was instrumental to effect differential deletions within complex hybrid transposons inserted in the chromosome of Pseudomonas putida. This strategy permits the stable inheritance of heterologous DNA segments virtually devoid of the sequences used initially to select their insertion.

M3 - Journal article

C2 - 7798149

VL - 177

SP - 52

EP - 58

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 1

ER -

ID: 44294108