[Protection of foreign DNA against host-controlled restriction in bacterial cells. II. Protection of pSF2124 plasmid by the gene function of bacteriophages T3 and T7].
When restriction-active Escherichia coli cells (R+P1m+P1) are transformed with the pSF2124 plasmid, a common vector in experimental gene transfer, the efficiency of transformation (e.o.t.) is lowered by 2 orders of magnitude compared with restriction-negative (r-P1m-P1 or r-P1m+P1) recipient cells due to restriction of the pSF2124 DNA by endoR.EcoP1. Preinfection of r+P1m+P1 cells with UV-inactivated ocr+ phages (T3, T7, T3sam-) still able to express their early genes protects the plasmid DNA against restriction by endoR.EcoP1: The e.o.t. of r+P1m+P1 recipient cells with pSF2124 attains the same high value as that of r-m- cells. The specific role of the ocr+ gene function was demonstrated by the use of ocr- mutants (T3/R7, T7/D111): Preinfection with such phage mutants does not increase the e.o.t. of r+P1m+P1 cells. An unspecific e.o.t. alteration of restriction-negative (r-m-) recipient cells by ocr+ or ocr- phages was excluded. The ocr+ gene function can be exploited to protect pSF2124 against DNA restriction. The recipient cells survive the process of phage preinfection and transformation and stably replicate themselves as well as the plasmid DNA.
D004262:DNA Restriction Enzymes / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005814:Genes, Viral; D005822:Genetic Vectors; D010957:Plasmids; D013604:T-Phages / Q000235:genetics* / Q000378:metabolism / Q000528:radiation effects; D014170:Transformation, Genetic; D014466:Ultraviolet Rays
D004279:DNA, Viral; D004262:DNA Restriction Enzymes
Schutz zellfremder DNA vor wirtskontrollierter Restriktion in Bakterienzellen. II. Schutz des Plasmids pSF2124 durch die ocr+ Genfunktion der Bakteriophagen T3 und T7.