A defined system for in vitro lambda DNA packaging.
We constructed a defined in vitro system for packaging lambda DNA which is composed of purified proheads, the noncapsid proteins terminase and gpFI, and the Escherichia coli DNA binding/bending protein IHF. The defined packaging system: (i) is free from endogenous ATP, DNAs, and DNases and (ii) packages 30% of the input mature lambda DNA efficiently. In this defined packaging system, IHF and gpFI make modest contributions to packaging efficiency. The defined packaging reactions showed that DNA packaging gave a linear response to the concentration of mature lambda DNA and terminase. DNA packaging showed a sigmoidal relationship with respect to the concentration of ATP and proheads.
Hwang
Y
Y
Department of Microbiology, University of Iowa, Iowa City 52242, USA.
Feiss
M
M
D000255:Adenosine Triphosphate / Q000494:pharmacology; D001426:Bacterial Proteins / Q000378:metabolism; D010582:Bacteriophage lambda / Q000235:genetics*; D004279:DNA, Viral / Q000187:drug effects / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000378:metabolism; D034982:Integration Host Factors; D014764:Viral Proteins / Q000378:metabolism
D001426:Bacterial Proteins; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D034982:Integration Host Factors; D014764:Viral Proteins; D000255:Adenosine Triphosphate; D004706:Endodeoxyribonucleases; C030894:terminase