Fluorescence label studies of the phase transitions of T7

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Abstract
Optical density, fluorescence intensity, polarization and lifetime measurements were performed to analyze the temperature-induced phase transitions of phage T7 in several buffers. For labelling the intraphage DNA, ethidium bromide, proflavine and rivanol were applied for the proteins 1,6-diphenyl-1,3,5-hexatriene (DPH). In the temperature range of 20‡C–100‡C several structural changes of T7 were detected. Making corrections for light scattering by using integrating spheres and analyzing the fluorescence signals, the phase transition of intraphage DNA below 60‡ C was interpreted as a superhelical relaxation phenomenon. The structural changes found by optical density at higher temperatures could be assigned to a change in the phage DNA or to a change in its protein part on the basis of fluorescence-melting results concerning DNA and protein labels. The effects of ionic strength and environment on the structural changes were studied.