EcoK restriction during in vitro packaging of coliphage lambda DNA.
The K restriction system of Escherichia coli works in vitro [Meselson and Yuan, Nature 217 (1968) 1110-1114]. E. coli C lacks the K restriction system. I show that in vitro packaging in standard E. coli K-12-derived systems effects a loss of plaque-former output from K-unmodified lambda DNA relative to K-modified lambda DNA when compared with packaging in the E. coli C-derived system of Rosenberg et al. [Gene 38 (1985) 165-175]. I conclude that the EcoK restriction system is active in standard in vitro packaging systems. EcoK restriction during in vitro packaging could specifically depress recovery of some lambda and cosmid clones of eukaryotic DNA or any other DNA not modified for EcoK restriction.
D010582:Bacteriophage lambda / Q000648:ultrastructure*; D002474:Cell-Free System; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D015253:Deoxyribonucleases, Type I Site-Specific; D009024:Morphogenesis
D004279:DNA, Viral; D004262:DNA Restriction Enzymes; C029602:endodeoxyribonuclease EcoK; D015253:Deoxyribonucleases, Type I Site-Specific