High-frequency transfer of linear DNA containing 5'-covalently linked terminal proteins: electroporation of bacteriophage PRD1 genome into Escherichia coli.
Using electroporation with the phage PRD1 genome, we set up a high-frequency DNA transfer system for a linear dsDNA molecule with 5'-covalently linked terminal proteins. The transfer was saturated when more than 100 ng of PRD1 genome was used. Electroporation efficiency was about four orders of magnitude higher than that obtained with transfection. Removal of the terminal protein abolished plaque formation, which could not be rescued by supplying the terminal protein or phage DNA polymerase or both in trans.
Lyra
C
C
Department of Genetics, University of Helsinki, Finland.
Savilahti
H
H
Bamford
D H
DH
D001435:Bacteriophages / Q000235:genetics*; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics; D005819:Genetic Markers; D010957:Plasmids; D014162:Transfection
D004274:DNA, Recombinant; D004279:DNA, Viral; D005819:Genetic Markers