Protein‐mediated DNA transfer into liposomes

Protein‐mediated DNA transfer into liposomes The transfer of a foreign genome into a bacterium by means of phage infection is a very efficient but poorly understood process. To analyse the mechanism of phage DNA transfer at a molecular level, we have reconstituted FhuA, the receptor for phage T5 in the outer membrane of Escherichia coli, into unilamellar vesicles made of natural phospholipids. Cryoelectron microscopy studies showed that the binding of the phage to FhuA triggered the transfer of its double‐stranded DNA (121 000 bp) into the proteoliposomes. DNA was entrapped within vesicles with a diameter ranging from 70 to 150 nm. The DNA appeared to be densely packed, but its presence did not alter the morphology of the liposomes, suggesting no DNA–lipid interactions. These liposomes represent an attractive model system for studying the mechanisms of DNA transport and condensation. They may also serve as alternative vehicles for the transfer of foreign genes into eukaryotic cells. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Molecular Microbiology Wiley

Protein‐mediated DNA transfer into liposomes

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Publisher
Wiley
Copyright
Blackwell Science Ltd, Oxford
ISSN
0950-382X
eISSN
1365-2958
DOI
10.1046/j.1365-2958.1998.01107.x
Publisher site
See Article on Publisher Site

Abstract

The transfer of a foreign genome into a bacterium by means of phage infection is a very efficient but poorly understood process. To analyse the mechanism of phage DNA transfer at a molecular level, we have reconstituted FhuA, the receptor for phage T5 in the outer membrane of Escherichia coli, into unilamellar vesicles made of natural phospholipids. Cryoelectron microscopy studies showed that the binding of the phage to FhuA triggered the transfer of its double‐stranded DNA (121 000 bp) into the proteoliposomes. DNA was entrapped within vesicles with a diameter ranging from 70 to 150 nm. The DNA appeared to be densely packed, but its presence did not alter the morphology of the liposomes, suggesting no DNA–lipid interactions. These liposomes represent an attractive model system for studying the mechanisms of DNA transport and condensation. They may also serve as alternative vehicles for the transfer of foreign genes into eukaryotic cells.

Journal

Molecular MicrobiologyWiley

Published: Nov 1, 1998

References

  • Cryo‐electron microscopy of vitrified specimens
    Dubochet, Dubochet; Adrian, Adrian; Chang, Chang; Hosuo, Hosuo; Lepault, Lepault; McDowall, McDowall
  • TonB‐dependent iron acquisition: mechanisms of siderophore‐mediated active transport
    Moeck, Moeck; Coulton, Coulton

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