Cristin-resultat-ID: 1121288
Sist endret: 30. oktober 2017, 11:04
NVI-rapporteringsår: 2014
Resultat
Vitenskapelig artikkel
2014

A new and improved host-independent plasmid system for RK2-based conjugal transfer

Bidragsytere:
  • Trine Aakvik
  • Rahmi Lale
  • Kristin Fløgstad Degnes
  • Malin Lando og
  • Svein Valla

Tidsskrift

PLOS ONE
ISSN 1932-6203
e-ISSN 1932-6203
NVI-nivå 1

Om resultatet

Vitenskapelig artikkel
Publiseringsår: 2014
Volum: 9
Hefte: 3:e90372
Open Access

Importkilder

Scopus-ID: 2-s2.0-84896925858

Beskrivelse Beskrivelse

Tittel

A new and improved host-independent plasmid system for RK2-based conjugal transfer

Sammendrag

Bacterial conjugation is a process that is mediated either by a direct cell-to-cell junction or by formation of a bridge between the cells. It is often used to transfer DNA constructs designed in Escherichia coli to recipient bacteria, yeast, plants and mammalian cells. Plasmids bearing the RK2/RP4 origin of transfer (oriT) are mostly mobilized using the E. coli S17-1/SM10 donor strains, in which transfer helper functions are provided from a chromosomally integrated RP4::Mu. We have observed that large plasmids were occasionally modified after conjugal transfer when using E. coli S17-1 as a donor. All modified plasmids had increased in size, which most probably was a result of co-transfer of DNA from the chromosomally located oriT. It has earlier also been demonstrated that the bacteriophage Mu is silently transferred to recipient cells by these donor strains, and both occurrences are very likely to lead to mutations within the recipient DNA. Here we report the construction of a new biological system addressing both the above mentioned problems in which the transfer helper functions are provided by a plasmid lacking a functional oriT. This system is compatible with all other replicons commonly used in conjugation experiments and further enables the use of diverse bacterial strains as donors. Plasmids containing large inserts were successfully conjugated and the plasmid modifications observed when E. coli S17-1 was used as donor were eliminated by the use of the new host-independent vector system.

Bidragsytere

Trine Aakvik Strand

Bidragsyterens navn vises på dette resultatet som Trine Aakvik
  • Tilknyttet:
    Forfatter
    ved Institutt for bioteknologi og matvitenskap ved Norges teknisk-naturvitenskapelige universitet

Rahmi Lale

  • Tilknyttet:
    Forfatter
    ved Institutt for bioteknologi og matvitenskap ved Norges teknisk-naturvitenskapelige universitet

Kristin Fløgstad Degnes

  • Tilknyttet:
    Forfatter
    ved Bioteknologi og nanomedisin ved SINTEF AS

Malin Lando

  • Tilknyttet:
    Forfatter
    ved Institutt for bioteknologi og matvitenskap ved Norges teknisk-naturvitenskapelige universitet

Svein Valla

  • Tilknyttet:
    Forfatter
    ved Institutt for bioteknologi og matvitenskap ved Norges teknisk-naturvitenskapelige universitet
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